Substituted pyridines and pyridazines with angiogenesis inhibiting activity

ABSTRACT

Substituted pyridazines having angiogenesis inhibiting activity and the generalized structural formula                    
     wherein the ring containing A, B, D, E, and L is phenyl or a nitrogen-containing heterocycle; groups X and Y may be any of a variety of defined linking units; R 1  and R 2  may be defined independent substituents or together may be a ring-defining bridge; ring J may be an aryl, pyridyl, or cycloalkyl group; and G groups may be any of a variety of defined substituents. Pharmaceutical compositions containing these materials, and methods of treating a mammal having a condition characterized by abnormal angiogenesis or hyperpermiability processes using these materials are also disclosed.

This application claims the benefit of U.S. Provisional Application No.60/287,595, filed Sep. 28, 1999.

FIELD

This application relates to small molecule heterocyclic pharmaceuticals,and more particularly, to substituted pyridines and pyridazines havingangiogenesis inhibiting activity.

BACKGROUND

Vasculogenesis involves the de novo formation of blood vessels fromendothelial cell precursors or angioblasts. The first vascularstructures in the embryo are formed by vasculogenesis. Angiogenesisinvolves the development of capillaries from existing blood vessels, andis the principle mechanism by which organs, such as the brain and thekidney are vascularized. While vasculogenesis is restricted to embryonicdevelopment, angiogenesis can occur in the adult, for example duringpregnancy, the female cycle, or wound healing.

One major regulator of angiogenesis and vasculogenesis in both embryonicdevelopment and some angiogenic-dependent diseases is vascularendothelial growth factor (VEGF; also called vascular permeabilityfactor, VPF). VEGF represents a family of mitogens isoforms resultingfrom alternative mRNA splicing and which exist in homodimeric forms. TheVEGF KDR receptor is highly specific for vascular endothelial cells (forreviews, see: Farrara et al. Endocr. Rev. 1992, 13, 18; Neufield et al.FASEB J. 1999, 13, 9).

VEGF expression is induced by hypoxia (Shweiki et al. Nature 1992, 359,843), as well as by a variety of cytokines and growth factors, such asinterleukin-1, interleukin-6, epidermal growth factor and transforminggrowth factor-α and -β.

To date VEGF and the VEGF family members have been reported to bind toone or more of three transmembrane receptor tyrosine kinases (Mustonenet al. J. Cell Biol., 1995, 129, 895), VEGF receptor-1 (also known asflt-1 (fms-like tyrosine kinase-1)); VEGFR-2 (also known as kinaseinsert domain containing receptor (KDR), the murine analogue of KDRbeing known as fetal liver kinase-1 (flk-1)); and VEGFR-3 (also known asflt-4). KDR and flt-1 have been shown to have different signaltransduction properties (Waltenberger et al. J. Biol. Chem. 1994, 269,26988); Park et al. Oncogene 1995, 10, 135). Thus, KDR undergoes strongligand-dependent tyrosine phosphorylation in intact cells, whereas flt-1displays a weaker response. Thus, binding to KDR is a criticalrequirement for induction of the full spectrum of VEGF-mediatedbiological responses.

In vivo, VEGF plays a central role in vasculogenesis, and inducesangiogenesis and permeabilization of blood vessels. Deregulated VEGFexpression contributes to the development of a number of diseases thatare characterized by abnormal angiogenesis and/or hyperpermeabilityprocesses. Regulation of the VEGF-mediated signal transduction cascadewill therefore provide a useful mode for control of abnormalangiogenesis and/or hyperpermeability processes.

Angiogenesis is regarded as an absolute prerequisite for growth oftumors beyond about 1-2 mm. Oxygen and nutrients may be supplied tocells in tumors smaller than this limit through diffusion. However,every tumor is dependent on angiogenesis for continued growth after ithas reached a certain size. Tumorigenic cells within hypoxic regions oftumors respond by stimulation of VEGF production, which triggersactivation of quiescent endothelial cells to stimulate new blood vesselformation. (Shweiki et al. Proc. Nat'l. Acad. Sci., 1995, 92, 768). Inaddition, VEGF production in tumor regions where there is noangiogenesis may proceed through the ras signal transduction pathway(Grugel et al. J. Biol. Chem., 1995, 270, 25915; Rak et al. Cancer Res.1995, 55, 4575). In situ hybridization studies have demonstrated VEGFmRNA is strongly upregulated in a wide variety of human tumors,including lung (Mattern et al. Br. J. Cancer 1996, 73, 931), thyroid(Viglietto et al. Oncogene 1995, 11, 1569), breast (Brown et al. HumanPathol. 1995, 26, 86), gastrointestional tract (Brown et al. Cancer Res.1993, 53, 4727; Suzuki et al. Cancer Res. 1996, 56, 3004), kidney andbladder (Brown et al. Am. J. Pathol. 1993, 143I, 1255), ovary (Olson etal. Cancer Res. 1994, 54, 1255), and cervical (Guidi et al. J. Nat'lCancer Inst. 1995, 87, 12137) carcinomas, as well as angiosacroma(Hashimoto et al. Lab. Invest. 1995, 73, 859) and several intracranialtumors (Plate et al. Nature 1992, 359, 845; Phillips et al. Int. J.Oncol. 1993, 2, 913; Berkman et al. J. Clin. Invest., 1993, 91, 153).Neutralizing monoclonal antibodies to KDR have been shown to beefficacious in blocking tumor angiogenesis (Kim et al. Nature 1993, 362,841; Rockwell et al. Mol. Cell. Differ. 1995, 3, 315).

Overexpression of VEGF, for example under conditions of extreme hypoxia,can lead to intraocular angiogenesis, resulting in hyperproliferation ofblood vessels, leading eventually to blindness. Such a cascade of eventshas been observed for a number of retinopathies, including diabeticretinopathy, ischemic retinal-vein occlusion, retinopathy of prematurity(Aiello et al. New Engl. J. Med. 1994, 331, 1480; Peer et al. Lab.Invest. 1995, 72, 638), and age-related macular degeneration (AMD; see,Lopez et al. Invest. Opththalmol. Vis. Sci. 1996, 37, 855).

In rheumatoid arthritis (RA), the in-growth of vascular pannus may bemediated by production of angiogenic factors. Levels of immunoreactiveVEGF are high in the synovial fluid of RA patients, while VEGF levelswere low in the synovial fluid of patients with other forms of arthritisof with degenerative joint disease (Koch et al. J. Immunol. 1994, 152,4149). The angiogenesis inhibitor AGM-170 has been shown to preventneovascularization of the joint in the rat collagen arthritis model(Peacock et al. J. Exper. Med. 1992, 175, 1135).

Increased VEGF expression has also been shown in psoriatic skin, as wellas bullous disorders associated with subepidermal blister formation,such as bullous pemphigoid, erythema multiforme, and dermatitisherpetiformis (Brown et al. J. Invest. Dermatol. 1995, 104, 744).

Because inhibition of KDR signal transduction leads to inhibition ofVEGF-mediated angiogenesis and permeabilization, KDR inhibitors will beuseful in treatment of diseases characterized by abnormal angiogenesisand/or hyperpermeability processes, including the above listed diseases.

Examples of phthalazines and other fused pyridazines that are similar instructure to those of the present application are disclosed in thefollowing patents or patent applications: WO 9835958 (Novartis), U.S.Pat. Nos. 5,849,741, 3,753,988, 3,478,028 and JP 03106875. Otherliterature references to phthalazines are El-Feky, S. A., Bayoumy, B.E., and Abd El-Sami, Z. K., Egypt. J. Chem. (1991), Volume Date 1990,33(2), 189-197; Duhault, J., Gonnard, P., and Fenard, S., Bull. Soc.Chim. Biol., (1967), 49 (2), 177-190; and Holava, H. M. and Jr, Partyka,R. A., J. Med. Chem., (1969), 12, 555-556. The compounds of the presentinvention are distinct from those described in each of the abovereferences, and only the Novartis publication describes such compoundsas inhibitors of angiogenesis.

As explained above, compounds which inhibit angiogenesis haveapplicability in treatment of a variety of medical conditions, and aretherefore desirable. Such materials are the subject of the presentapplication.

SUMMARY

In its broadest aspect, the present invention relates to the sum ofthree sets of chemical compounds, or pharmaceutically acceptable saltsor prodrugs thereof, with each set overlapping the others in scope. Thegeneralized structural formula for the compounds in each of the threesets of compounds is the same, but it should be noted that thedefinitions of the several groups comprising the general structure ineach set differ somewhat. Thus, the defined sets of chemical compoundsdiffer from each other, but overlap in their scopes.

The first set of compounds have the generalized structural formula

wherein

R¹ and R² together form a bridge containing two T² moieties and one T³moiety, said bridge, taken together with the ring to which it isattached, forming a bicyclic of structure

 wherein

each T² independently represents N, CH, or CG¹;

T³ represents S, O, CR⁴G¹, C(R⁴)₂, or NR³.

In the above substructures, G¹ is a substituent independently selectedfrom the group consisting of —N(R⁶)₂; —NR³COR⁶; halogen; alkyl;cycloalkyl; lower alkenyl; lower cycloalkenyl; halogen-substitutedalkyl; amino-substituted alkyl; N-lower alkylamino-substituted alkyl;N,N-di-lower alkylamino-substituted alkyl; N-loweralkanoylamino-substituted alkyl; hydroxy-substituted alkyl;cyano-substituted alkyl; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted saturated heterocyclylalkyl;optionally substituted partially unsaturated heterocyclyl; optionallysubstituted partially unsaturated heterocyclylalkyl; —OCO₂R³; optionallysubstituted heteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; and —NR³CON(R⁶)₂.

The group R³ is H or lower alkyl. R⁶ is independently selected from thegroup consisting of H; alkyl; cycloalkyl; optionally substituted aryl;optionally substituted aryl lower alkyl, lower alkyl-N(R³)₂, and loweralkyl-OH.

In generalized structural formula (I), R⁴ is H, halogen, or lower alkyl.The subscript p is 0, 1, or 2; and X is selected from the groupconsisting of O, S, and NR³.

The linking moiety Y is selected from the group consisting of loweralkylene; —CH₂—O—; —CH₂—S—; —CH₂—NH—; —O—; —S—; —NH—; —O—CH₂—; —S(O)—;—S(O)₂—; —SCH₂—; —S(O)CH₂—; —S(O)₂CH₂—; —CH₂S(O)—; —CH₂S(O)₂; —(CR⁴₂)_(n)—S(O)_(p)-(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; and —(CR⁴₂)_(n)—C(G²)(R⁴)—(CR⁴ ₂)_(s)—. In the latter two linking groups Y, n ands are each independently 0 or an integer of 1-2. The substituent G² isselected from the group consisting of —CN, —CO₂R³, —CON(R⁶)₂, and—CH₂N(R⁶)₂.

Z represents CR⁴ or N.

Regarding the ring containing A, B, D, E, and L, the number of possiblesubstituents G³ on the ring is indicated by subscript q, which is 0, 1,or 2.

Substituent moieties G³ are monovalent or bivalent moieties selectedfrom the group consisting of: lower alkyl; —NR³COR⁶; carboxy-substitutedalkyl; lower alkoxycarbonyl-substituted alkyl; —OR⁶; —SR⁶; —S(O)R⁶;—S(O)₂R⁶; —OCOR⁶; —COR⁶; —CO₂R⁶; —CH₂OR³; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; —NO₂;—CN; optionally substituted aryl; optionally substituted heteroaryl;optionally substituted saturated heterocyclyl; optionally substitutedpartially unsaturated heterocyclyl; optionally substitutedheteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂; and bivalent bridge of structureT²═T²—T³. In this bivalent bridge, each T² independently represents N,CH, or CG^(3′); and T³ represents S, O, CR⁴G^(3′), C(R⁴)₂, or NR³.G^(3′) represents any of the above-defined moieties G³ which aremonovalent; and the terminal T² of the bridge is bound to L, and T³ isbound to D, thus forming a 5-membered fused ring.

In the ring shown at the left in generalized structural formula (I), Aand D independently represent N or CH; B and E independently represent Nor CH; and L represents N or CH; with the provisos that a) the totalnumber of N atoms in the ring containing A, B, D, E, and L is 0, 1, 2,or 3; b) when L represents CH and any G³ is a monovalent substituent, atleast one of A and D is an N atom; and c) when L represents CH and a G³is a bivalent bridge of structure T²═T²—T³, then A, B, D, and E are alsoCH.

J is a ring selected from the group consisting of aryl; pyridyl; andcycloalkyl. The subscript q′ represents the number of substituents G⁴ onring J and is 0, 1, 2, 3, 4, or 5.

The possible substituents G⁴ on ring J are monovalent or bivalentmoieties selected from the group consisting of —N(R⁶)₂; —NR³COR⁶;halogen; alkyl; cycloalkyl; lower alkenyl; lower cycloalkenyl;halogen-substituted alkyl; amino-substituted alkyl; N-loweralkylamino-substituted alkyl; N,N-di-lower alkylamino-substituted alkyl;N-lower alkanoylamino-substituted alkyl; hydroxy-substituted alkyl;cyano-substituted alkyl; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted partially unsaturated heterocyclyl;—OCO₂R³; optionally substituted heteroarylalkyl; optionally substitutedheteroaryloxy; —S(O)_(p)(optionally substituted heteroaryl); optionallysubstituted heteroarylalkyloxy; —S(O)_(p)(optionally substitutedheteroarylalkyl); —CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)2; and fusedring-forming bivalent bridges attached to and connecting adjacentpositions of ring J, said bridges having the structures:

 wherein each T² independently represents N, CH, or CG^(4′); T³represents S, O, CR⁴G^(4′), C(R⁴)₂, or NR³; G4′ represents any of theabove-defined moieties G⁴ which are monovalent; and binding to ring J isachieved via terminal atoms T² and T³;

 wherein each T² independently represents N, CH, or CG^(4′); G4′represents any of the above-defined moieties G⁴ which are monovalent;with the proviso that a maximum of two bridge atoms T² may be N; andbinding to ring J is achieved via terminal atoms T²; and

 wherein each T⁴, T⁵ and T⁶ independently represents O, S, CR⁴G^(4′),C(R⁴)₂, or NR³; G4′ represents any of the above-defined moieties G⁴which are monovalent; and binding to ring J is achieved via terminalatoms T⁴ or T⁵; with the provisos that:

i) when one T⁴ is O, S, or NR³, the other T⁴ is CR⁴G^(4′) or C(R⁴)₂;

ii) a bridge comprising T⁵ and T⁶ atoms may contain a maximum of twoheteroatoms O, S, or N; and

iii) in a bridge comprising T⁵ and T⁶ atoms, when one T⁵ group and oneT⁶ group are O atoms, or two T⁶ groups are O atoms, said O atoms areseparated by at least one carbon atom.

When G⁴ is an alkyl group located on ring J adjacent to the linkage—(CR⁴ ₂)_(p)—, and X is NR³ wherein R³ is an alkyl substituent; then G⁴and the alkyl substituent R³ on X may be joined to form a bridge ofstructure —(CH₂)_(p′)— wherein p′ is 2, 3, or 4, with the proviso thatthe sum of p and p′ is 2, 3, or 4, resulting in formation of anitrogen-containing ring of 5, 6, or 7 members.

Additional provisos are that: 1) in G¹, G², G³, and G⁴, when two groupsR³ or R⁶ are each alkyl and located on the same N atom they may belinked by a bond, an O, an S, or NR³ to form a N-containing heterocycleof 5-7 ring atoms; and 2) when an aryl, heteroaryl, or heterocyclyl ringis optionally substituted, that ring may bear up to 5 substituents whichare independently selected from the group consisting of amino,mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino,lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl,hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy,halogenated lower alkylthio, lower alkanoyloxy, —CO₂R³, —CHO, —CH₂OR³,—OCO₂R³, —CON(R⁶)₂, —OCON(R⁶)₂, —NR³CON(R⁶)₂, nitro, amidino, guanidino,mercapto, sulfo, and cyano; and 3) when any alkyl group is attached toO, S, or N, and bears a hydroxyl substituent, then said hydroxylsubstituent is separated by at least two carbon atoms from the O, S, orN to which the alkyl group is attached.

The second set of compounds have the generalized structural formula

wherein

R¹ and R²:

i) independently represent H or lower alkyl;

ii) together form a bridge of structure

 wherein binding is achieved via the terminal carbon atoms;

iii) together form a bridge of structure

 wherein binding is achieved via the terminal carbon atoms;

iv) together form a bridge of structure

 wherein one or two ring members T¹ are N and the others are CH or CG¹,and binding is achieved via the terminal atoms; or

v) together form a bridge containing two T² moieties and one T³ moiety,said bridge, taken together with the ring to which it is attached,forming a bicyclic of structure

 wherein

each T² independently represents N, CH, or CG¹;

T³ represents S, O, CR⁴G¹, C(R⁴)₂, or NR³.

In the above bridge substructures, the subscript m is 0 or an integer1-4; indicating that the resultant fused rings may optionally bear up tofour substituents G¹.

G¹ is a substituent independently selected from the group consisting of—N(R⁶)₂; —NR³COR⁶; halogen; alkyl; cycloalkyl; lower alkenyl; lowercycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl;N-lower alkylamino-substituted alkyl; N,N-di-loweralkylamino-substituted alkyl; N-lower alkanoylamino-substituted alkyl;hydroxy-substituted alkyl; cyano-substituted alkyl; carboxy-substitutedalkyl; lower alkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted saturated heterocyclylalkyl;optionally substituted partially unsaturated heterocyclyl; optionallysubstituted partially unsaturated heterocyclylalkyl; —OCO₂R³; optionallysubstituted heteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; and —NR³CON(R⁶)₂.

The group R³ is H or lower alkyl. R⁶ is independently selected from thegroup consisting of H; alkyl; cycloalkyl; optionally substituted aryl;optionally substituted aryl lower alkyl; lower alkyl-N(R³)₂, and loweralkyl-OH.

In generalized structural formula (I), R⁴ is H, halogen, or lower alkyl;the subscript p is 0, 1, or 2; and X is selected from the groupconsisting of O, S, and NR³.

The linking moiety Y is selected from the group consisting of loweralkylene; —CH₂O—; —CH₂—S—; —CH₂—NH—; —O—; —S—; —NH—; —O—CH₂—; —S(O)—;—S(O)₂—; —SCH₂—; —S(O)CH₂—; —S(O)₂CH₂—; —CH₂S(O)—; —CH₂S(O)₂—; —(CR⁴₂)_(n)—S(O)_(p)—(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; and —(CR⁴₂)_(n)—C(G²)(R⁴)—(CR⁴ ₂)_(s)—. In the latter two linking groups Y,subscripts n and s are each independently 0 or an integer of 1-2. G² isselected from the group consisting of —CN, —CO₂R³, —CON(R⁶)₂, and—CH₂N(R⁶)₂.

Z represents N or CR⁴.

Regarding the ring containing A, B, D, E, and L, the number of possiblesubstituents G³ on the ring is indicated by the subscript q, which is 1or 2.

Substituents G³ are monovalent or bivalent moieties selected from thegroup consisting of lower alkyl; —NR³COR⁶; carboxy-substituted alkyl;lower alkoxycarbonyl-substituted alkyl; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;—OCOR⁶; —COR⁶; —CO₂R⁶; —CH₂OR³; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; —NO₂; —CN;optionally substituted aryl; optionally substituted heteroaryl;optionally substituted saturated heterocyclyl; optionally substitutedpartially unsaturated heterocyclyl; optionally substitutedheteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂; and bivalent bridge of structureT²═T²—T³. In this bivalent bridge, each T² independently represents N,CH, or CG^(3′); and T³ represents S, O, CR⁴G^(3′), C(R⁴)₂, or NR³. G³represents any of the above-defined moieties G³ which are monovalent;and the terminal T² is bound to L, and T³ is bound to D, thus forming a5-membered fused ring.

In the ring shown at the left in generalized structural formula (I), Aand D independently represent CH; B and E independently represent CH;and L is CH; with the proviso that the resulting phenyl ring bears as aG³ substituent said bivalent bridge of structure T²═T²—T³.

J is a ring selected from the group consisting of aryl; pyridyl; andcycloalkyl. The subscript q′ represents the number of substituents G⁴ onring J and is 0, 1, 2, 3, 4, or 5.

G⁴ is a monovalent or bivalent moiety selected from the group consistingof —N(R⁶)₂; —NR³COR⁶; halogen; alkyl; cycloalkyl; lower alkenyl; lowercycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl;N-lower alkylamino-substituted alkyl; N,N-di-loweralkylamino-substituted alkyl; N-lower alkanoylamino-substituted alkyl;hydroxy-substituted alkyl; cyano-substituted alkyl; carboxy-substitutedalkyl; lower alkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted partially unsaturated heterocyclyl;—OCO₂R³; optionally substituted heteroarylalkyl; optionally substitutedheteroaryloxy; —S(O)_(p)(optionally substituted heteroaryl); optionallysubstituted heteroarylalkyloxy; —S(O)_(p)(optionally substitutedheteroarylalkyl); —CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂; and fusedring-forming bivalent bridges attached to and connecting adjacentpositions of ring J, said bridges having the structures:

 wherein each T² independently represents N, CH, or CG^(4′); T³represents S, O, CR⁴G^(4′), C(R⁴)₂, or NR³; G^(4′) represents any of theabove-defined moieties G⁴ which are monovalent; and binding to ring J isachieved via terminal atoms T² and T³;

 wherein each T² independently represents N, CH, or CG^(4′); G4′represents any of the above-defined moieties G⁴ which are monovalent;with the proviso that a maximum of two bridge atoms T² may be N; andbinding to ring J is achieved via terminal atoms T²; and

 wherein each T⁴, T⁵, and T⁶ independently represents O, S, CR⁴G^(4′),C(R⁴)₂, or NR³; G4′ represents any of the above-identified moieties G⁴which are monovalent; and binding to ring J is achieved via terminalatoms T⁴ or T⁵; with the provisos that:

i) when one T⁴ is O, S, or NR³, the other T⁴ is CR⁴G^(4′) or C(R⁴)₂;

ii) a bridge comprising T⁵ and T⁶ atoms may contain a maximum of twoheteroatoms O, S, or N; and

iii) in a bridge comprising T⁵ and T⁶ atoms, when one T⁵ group and oneT⁶ group are O atoms, or two T⁶ groups are O atoms, said O atoms areseparated by at least one carbon atom.

When G⁴ is an alkyl group located on ring J adjacent to the linkage —CR⁴₂)_(p)—, and X is NR³ wherein R³ is an alkyl substituent, then G⁴ andthe alkyl substituent R³ on X may be joined to form a bridge ofstructure —(CH₂)_(p′)— wherein p′ is 2, 3, or 4, with the proviso thatthe sum of p and p′ is 2, 3, or 4, resulting in formation of anitrogen-containing ring of 5, 6, or 7 members.

Additional provisos are that: 1) in G¹, G², G³, and G⁴, when two groupsR³ or R⁶ are each alkyl and located on the same N atom they may belinked by a bond, an O, an S, or NR³ to form a N-containing heterocycleof 5-7 ring atoms; and 2) when an aryl, heteroaryl, or heterocyclyl ringis optionally substituted, that ring may bear up to 5 substituents whichare independently selected from the group consisting of amino,mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino,lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl,hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy,halogenated lower alkylthio, lower alkanoyloxy, —CO₂R³, —CHO, —CH₂OR³,—OCO₂R³, —CON(R⁶)₂, —OCON(R⁶)₂, —NR³CON(R⁶)₂, nitro, amidino, guanidino,mercapto, sulfo, and cyano; and 3) when any alkyl group is attached toO, S, or N, and bears a hydroxyl substituent, then said hydroxylsubstituent is separated by at least two carbon atoms from the O, S, orN to which the alkyl group is attached.

The third set of compounds have the generalized structural formula

wherein

R¹ and R²:

i) independently represent H or lower alkyl;

ii) together form a bridge of structure

 wherein binding is achieved via the terminal carbon atoms;

iii) together form a bridge of structure

 wherein binding is achieved via the terminal carbon atoms;

iv) together form a bridge of structure

 wherein one or two ring members T¹ are N and the others are CH or CG¹,and binding is achieved via the terminal atoms; or

v) together form a bridge containing two T² moieties and one T³ moiety,said bridge, taken together with the ring to which it is attached,forming a bicyclic of structure

 wherein

each T² independently represents N, CH, or CG¹;

T³ represents S, O, CR⁴G¹, C(R⁴)₂, or NR³.

In the above bridge structures, the subscript m is 0 or an integer 1-4;indicating that the resultant fused rings may optionally bear up to foursubstituents G¹.

G¹ is a substituent independently selected from the group consisting of—N(R⁶)₂; —NR³COR⁶; halogen; alkyl; cycloalkyl; lower alkenyl; lowercycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl;N-lower alkylamino-substituted alkyl; N,N-di-loweralkylamino-substituted alkyl; N-lower alkanoylamino-substituted alkyl;hydroxy-substituted alkyl; cyano-substituted alkyl; carboxy-substitutedalkyl; lower alkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted saturated heterocyclylalkyl;optionally substituted partially unsaturated heterocyclyl; optionallysubstituted partially unsaturated heterocyclylalkyl; —OCO₂R³; optionallysubstituted heteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; and —NR³CON(R⁶)₂.

The group R³ is H or lower alkyl. R⁶ is independently selected from thegroup consisting of H; alkyl; cycloalkyl; optionally substituted aryl;optionally substituted aryl lower alkyl; lower alkyl-N(R³)₂, and loweralkyl-OH.

In generalized structural formula (I), R⁴ is H, halogen, or lower alkyl;the subscript p is 0, 1, or 2; and X is selected from the groupconsisting of O, S, and NR³.

The linking moiety Y is selected from the group consisting of loweralkylene; —CH₂O—; —CH₂—S—; —CH₂—NH—; —O—; —S—; —NH—; —O—CH₂—; —S(O)—;—S(O)₂—; —SCH₂—; —S(O)CH₂—; —S(O)₂CH₂—; —CH₂S(O)—; —CH₂S(O)₂—; —(CR⁴₂)_(n)—S(O)_(p)—(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; and —(CR⁴₂)_(n)—C(G²)(R⁴)—(CR⁴ ₂)_(s)—. In the latter two linking groups Y,subscripts n and s are each independently 0 or an integer of 1-2. G² isselected from the group consisting of —CN, —CO₂R³, —CON(R⁶)₂, and—CH₂N(R⁶)₂.

Z represents CR⁴.

Regarding the ring containing A, B, D, E, and L, the number of possiblesubstituents G³ on the ring is indicated by the subscript q, which is 1or 2.

Substituents G are monovalent or bivalent moieties selected from thegroup consisting of —NR³COR⁶; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶; —OCOR⁶;—COR⁶; —CO₂R⁶; —CH₂OR³; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; —NO₂; —CN; optionallysubstituted aryl; optionally substituted heteroaryl; optionallysubstituted saturated heterocyclyl; optionally substituted partiallyunsaturated heterocyclyl; optionally substituted heteroarylalkyl;optionally substituted heteroaryloxy; —S(O)_(p)(optionally substitutedheteroaryl); optionally substituted heteroarylalkyloxy;—S(O)_(p)(optionally substituted heteroarylalkyl); —OCON(R⁶)₂;—NR³CO₂R⁶; —NR³CON(R⁶)₂; and bivalent bridge of structure T²═T²—T³. Inthis bivalent bridge, each T² independently represents N, CH, orCG^(3′); and T³ represents S, O, CR⁴G^(3′), C(R⁴)₂, or NR³. G^(3′)represents any of the above-defined moieties G³ which are monovalent;and the terminal T² is bound to L, and T³ is bound to D, thus forming a5-membered fused ring.

In the ring shown at the left in generalized structural formula (I), Aand D independently represent N or CH; B and E independently represent Nor CH; and L represents N or CH; with the provisos that a) the totalnumber of N atoms in the ring containing A, B, D, E, and L is 0, 1, 2,or 3; and b) when L represents CH and any G³ is a monovalentsubstituent, at least one of A and D is an N atom; and c) when Lrepresents CH and a G³ is a bivalent bridge of structure T²═T²—T³, thenA, B, D, and E are also CH.

J is a ring selected from the group consisting of aryl; pyridyl; andcycloalkyl. The subscript q′ represents the number of substituents G⁴ onring J and is 0, 1, 2, 3, 4, or 5.

G⁴ is a monovalent or bivalent moiety selected from the group consistingof —N(R⁶)₂; —NR³COR⁶; halogen; alkyl; cycloalkyl; lower alkenyl; lowercycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl;N-lower alkylamino-substituted alkyl; N,N-di-loweralkylamino-substituted alkyl; N-lower alkanoylamino-substituted alkyl;hydroxy-substituted alkyl; cyano-substituted alkyl; carboxy-substitutedalkyl; lower alkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted partially unsaturated heterocyclyl;—OCO₂R³; optionally substituted heteroarylalkyl; optionally substitutedheteroaryloxy; —S(O)_(p)(optionally substituted heteroaryl); optionallysubstituted heteroarylalkyloxy; —S(O)_(p)(optionally substitutedheteroarylalkyl); —CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂; and fusedring-forming bivalent bridges attached to and connecting adjacentpositions of ring J, said bridges having the structures:

 wherein each T² independently represents N, CH, or CG^(4′); T³represents S, O, CR⁴G^(4′), C(R⁴)₂, or NR³; G4′ represents any of theabove-defined moieties G⁴ which are monovalent; and binding to ring J isachieved via terminal atoms T² and T³;

 wherein each T² independently represents N, CH, or CG^(4′); G4′represents any of the above-defined moieties G⁴ which are monovalent;with the proviso that a maximum of two bridge atoms T² may be N; andbinding to ring J is achieved via terminal atoms T²; and

 wherein each T⁴, T⁵, and T⁶ independently represents O, S, CR⁴G^(4′),C(R⁴)₂, or NR³; G4′ represents any of the above-defined moieties G⁴which are monovalent; and binding to ring J is achieved via terminalatoms T⁴ or T ⁵; with the provisos that:

i) when one T⁴ is O, S, or NR³, the other T is CR⁴G^(4′) or C(R⁴)₂;

ii) a bridge comprising T⁵ and T⁶ atoms may contain a maximum of twoheteroatoms O, S, or N; and

iii) in a bridge comprising T⁵ and T⁶ atoms, when one T⁵ group and oneT⁶ group are O atoms, or two T⁶ groups are O atoms, said O atoms areseparated by at least one carbon atom;

When G⁴ is an alkyl group located on ring J adjacent to the linkage—(CR⁴ ₂)_(p)—, and X is NR³ wherein R³ is an alkyl substituent, then G⁴and the alkyl substituent R³ on X may be joined to form a bridge ofstructure —(CH₂)_(p′)— wherein p′ is 2, 3, or 4, with the proviso thatthe sum of p and p′ is 2, 3, or 4, resulting in formation of anitrogen-containing ring of 5, 6, or 7 members.

Additional provisos are that: 1) in G¹, G², G³, and G⁴, when two groupsR³ or R⁶ are each alkyl and located on the same N atom they may belinked by a bond, an O, an S, or NR³ to form a N-containing heterocycleof 5-7 ring atoms; and 2) when an aryl, heteroaryl, or heterocyclyl ringis optionally substituted, that ring may bear up to 5 substituents whichare independently selected from the group consisting of amino,mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino,lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl,hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy,halogenated lower alkylthio, lower alkanoyloxy, —CO₂R³, —CHO, —CH₂OR³,—OCO₂R³, —CON(R⁶)₂, —OCON(R⁶)₂, —NR³CON(R⁶)₂, nitro, amidino, guanidino,mercapto, sulfo, and cyano; and 3) when any alkyl group is attached toO, S, or N, and bears a hydroxyl substituent, then said hydroxylsubstituent is separated by at least two carbon atoms from the O, S, orN to which the alkyl group is attached.

Pharmaceutically acceptable salts of these compounds as well as commonlyused prodrugs of these compounds such as O-acyl derivatives of inventioncompounds which contain hydroxy groups are also within the scope of theinvention.

The invention also relates to pharmaceutical compositions comprising oneor more of the compounds of the invention, or their salts or prodrugs,in a pharmaceutically acceptable carrier.

The invention also relates to a method for using these materials totreat a mammal having a condition characterized by abnormal angiogenesisor hyperpermiability processes, comprising administering to the mammalan amount of a compound of the invention, or a salt or prodrug thereof,which is effective to treat the condition.

DETAILED DESCRIPTION Definitions

The prefix “lower” denotes a radical having up to and including amaximum of 7 atoms, especially up to and including a maximum of 5 carbonatoms, the radicals in question being either linear or branched withsingle or multiple branching.

“Alkyl” means a hydrocarbon radical having up to a maximum of 12 carbonatoms, which may be linear or branched with single or multiplebranching. Alkyl is especially lower alkyl.

Where the plural form is used for compounds, salts, and the like, thisis taken to mean also a single compound, salt, or the like.

Any asymmetric carbon atoms may be present in the (R)-, (S)- or(R,S)configuration, preferably in the (R)- or (S)-configuration.Substituents at a double bond or a ring may be present in cis- (═Z-) ortrans (═E-) form. The compounds may thus be present as mixtures ofisomers or as pure isomers, preferably as enantiomer-pure diastereomersand having pure cis- or trans-double bonds.

Lower alkylene Y may be branched or linear but is preferably linear,especially methylene (—CH₂), ethylene (—CH₂—CH₂), trimethylene(—CH₂—CH₂—CH₂) or tetramethylene (—CH₂CH₂CH₂CH₂). When Y is loweralkylene, it is most preferably methylene.

“Aryl” means an aromatic radical having 6 to 14 carbon atoms, such asphenyl, naphthyl, fluorenyl or phenanthrenyl.

“Halogen” means fluorine, chlorine, bromine, or iodine but is especiallyfluorine, chlorine, or bromine.

“Pyridyl” means 1-, 2-, or 3-pyridyl but is especially 2- or 3-pyridyl.

“Cycloalkyl” is a saturated carbocycle that contains between 3 and 12carbons but preferably 3 to 8 carbons.

“Cycloalkenyl” means a non-reactive and non-aromatic unsaturatedcarbocycle that contains between 3 and 12 carbons but preferably 3 to 8carbons and up to three double bonds. It is well known to those skilledin the art that cycloalkenyl groups that differ from aromatics bylacking only one double bond such as cyclohaxadiene are not sufficientlynon-reactive to be reasonable drug substances and therefor their use assubstituents is not within the scope of this invention.

Cycloalkyl and cycloalkenyl groups may contain branch points such thatthey are substituted by alkyl or alkenyl groups. Examples of suchbranched cyclic groups are 3,4-dimethylcyclopentyl, 4-allylcyclohexyl or3-ethylcyclopent-3-enyl.

Salts are especially the pharmaceutically acceptable salts of compoundsof formula I such as, for example, acid addition salts, preferably withorganic or inorganic acids, from compounds of formula I with a basicnitrogen atom. Suitable inorganic acids are, for example, halogen acidssuch as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitableorganic acids are, for example, carboxylic, phosphonic, sulfonic, orsulfamic acids, for example acetic acid, propionic acid, octanoic acid,decanoic acid, dodecanoic acid, glycolic acid, lactic acid,-hydroxybutyric acid, gluconic acid, glucosemonocarboxylic acid, fumaricacid, succinic acid, adipic acid, pimelic acid, suberic acid, azeiaicacid, malic acid, tartaric acid, citric acid, glucaric acid, galactaricacid, amino acids, such as glutamic acid, aspartic acid,N-methylglycine, acetytaminoacetic acid, N-acetylasparagine orN-acetylcysteine, pyruvic acid, acetoacetic acid, phosphoserine, 2- or3-glycerophosphoric acid.

In the definition of Y, the diradical “-(5 member heteroaryl)-” denotesa 5-membered aromatic heterocycle containing 1-3 heteroatoms selectedfrom O, S, and N, the number of N atoms being 0-3 and the number of Oand S atoms each being 0-1 and connected to the sulfur from a carbon andto —(CR⁴ ₂)_(s)— through a C or N atom. Examples of such diradicalsinclude

In the definitions of G¹, G², G³, and G⁴ the statement is made that whentwo groups R³ or R⁶ are found on a single N, they can be combined into aheterocycle of 5-7 atoms. Examples of such heterocycles, including the Nto which they are attached, are:

“Heterocyclyl” or “heterocycle” means a five- to seven-memberedheterocyclic system with 1-3 heteroatoms selected from the groupnitrogen, oxygen, and sulfur, which may be unsaturated or wholly orpartly saturated, and is unsubstituted or substituted especially bylower alkyl, such as methyl, ethyl, 1-propyl, 2-propyl, or tert-butyl.

When an aryl, heteroaryl, or heterocyclyl ring is said to be optionallysubstituted, that ring may bear up to 5 substituents which areindependently selected from the group consisting of amino, mono- ordi-loweralkyl-substituted amino, lower alkanoylamino, halogeno, loweralkyl, halogenated lower alkyl such as trifluoromethyl, hydroxy, loweralkoxy, lower alkylthio, halogenated lower alkoxy such astrifluoromethoxy, halogenated lower alkylthio such astrifluoromethylthio, lower alkanoyloxy, —CO₂R³, —CHO, —CH₂OR³, —OCO₂R³,—CON(R⁶)₂, —OCON(R⁶)₂, —NR³CON(R⁶)₂, nitro, amidino, guanidino,mercapto, sulfo, and cyano.

In the ring attached to Y, the ring members A, B, D, E, and L may be Nor CH, it being understood that the optional substituents G³ arenecessarily attached to carbon and not nitrogen, and that when a givencarbon bears a substituent group G³, that G³ group is in place of the Hatom the carbon would bear in the absence of the G³ group.

Examples of ring J together with two adjacent G⁴ moieties which takentogether form a second fused ring are:

“Heteroaryl” means a monocyclic or fused bicyclic aromatic system withbetween 5 and 10 atoms in total of which 1-4 are heteroatoms selectedfrom the group comprising nitrogen, oxygen, and sulfur and with theremainder being carbon. Heteroaryl is preferably a monocyclic systemwith 5 or 6 atoms in total, of which 1-3 are heteroatoms.

“Alkenyl” means an unsaturated radical having up to a maximum of 12carbon atoms and may be linear or branched with single or multiplebranching and containing up to 3 double bonds. Alkenyl is especiallylower alkenyl with up to 2 double bonds.

“Alkanoyl” means alkylcarbonyl, and is especially lower alkylcarbonyl.

Halogenated lower alkyl, halogenated lower alkoxy and halogenated loweralkylthio are substituents in which the alkyl moieties are substitutedeither partially or in full with halogens, preferably with chlorineand/or fluorine and most preferably with fluorine. Examples of suchsubstituents are trifluoromethyl, trifluoromethoxy, trifluoromethylthio,1,1,2,2-tetrafluoroethoxy, dichloromethyl, fluoromethyl anddifluoromethyl.

When a substituent is named as a string of fragments such as“phenyl-lower alkoxycarbonyl-substituted alkylamino”, it is understoodthat the point of attachment is to the final moiety of that string (inthis case amino) and that the other fragments of that string areconnected to each other in sequence as they are listed in the string.Thus an example of “phenyl-lower alkoxycarbonyl-substituted alkylamino”is:

When a substituent is named as a string of fragments with a bond at thestart (typically written as a dash) such as “—S(O)_(p)(optionallysubstituted heteroarylalkyl)”, it is understood that the point ofattachment is to the first atom of that string (in this case S orsulfur) and that the other fragments of that string are connected toeach other in sequence as they are listed in the string. Thus an exampleof “—S(O)_(p)(optionally substituted heteroarylalkyl)” is:

It is to be understood that the left-most moiety of each of the variantsof the linker Y is connected to the ring containing A, B, D, E, and Land that the right-most moiety of the linker is connected to thepyridazine fragment of the generalized formulae. Thus, examples of theuse of the linker “—CH₂—O—” or of the linker “—O—CH₂—” are representedin the following invention compounds:

In generalized structural formula (I), the preferred and most preferredgroups are as follows.

R¹ and R² preferably:

i) together form a bridge of structure

 wherein binding is achieved via the terminal carbon atoms; or

ii) together form a bridge of structure

 wherein one of the ring members T¹ is N and the others are CH, andbinding is achieved via the terminal atoms; or

iii) together form a bridge containing two T² moieties and one T³moiety, said bridge, taken together with the ring to which it isattached, forming a bicyclic of structure

 wherein

each T² independently represents N, CH, or CG¹;

T³ represents S, O, CH₂, or NR³; and

with the proviso that when T³ is O or S, at least one T² is CH or CG¹.

Most preferably, any group G¹ is located on a non-terminal atom of thebridge. Most preferably, in the bridge in iii), the terminal T² is N orCH, the non-terminal T² is CH or CG¹, and T³ is S or O.

The subscript m is preferably 0 or an integer 1-2, and substituents G¹are preferably selected from the group consisting of —N(R⁶)₂; —NR³COR⁶;halogen; lower alkyl; hydroxy-substituted alkyl; amino-substitutedalkylamino; N-lower alkylamino-substituted alkylamino; N,N-di-loweralkylamino-substituted alkylamino; hydroxy-substituted alkylamino;carboxy-substituted alkylamino; lower alkoxycarbonyl-substitutedalkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶; halogenated lower alkoxy;halogenated lower alkylthio; halogenated lower alkylsulfonyl; —OCOR⁶;—COR⁶; —CO₂R⁶; —CON(R⁶)₂; —NO₂; —CN; optionally substitutedheteroarylalkyl; optionally substituted heteroaryloxy; optionallysubstituted heteroarylalkyloxy; and —S(O)_(p)(optionally substitutedheteroarylalkyl). Most preferably, m is 0, and G¹ is a substituentindependently selected from the group consisting of —N(R⁶)₂; —NR³COR⁶;halogen; —OR⁶ wherein R6 represents lower alkyl; —NO₂; optionallysubstituted heteroaryloxy; and optionally substitutedheteroarylalkyloxy.

When R⁶ is an alkyl group, it is preferably lower alkyl. The group R⁴ ispreferably H; p is preferably 0 or 1; and X is preferably NR³.

In the linker group Y, the subscripts n and s are preferably 0 or 1,most preferably 0. Preferably, Y is selected from the group consistingof lower alkylene, —CH₂—O—; —CH₂—S—; —CH₂—NH—; —S—; —NH—; —(CR⁴₂)_(n)—S(O)_(p)—(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; —(CR⁴₂)_(n)—C(G²)(R⁴)—(CR⁴ ₂)_(s)—; and —O—CH₂—. Most preferably, Y isselected from the group consisting of —CH₂—O—; —CH₂—NH—; —S—; —NH—;—(CR⁴ ₂)_(n)—S(O)_(p)—(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; and —O—CH₂—.

In the ring at the left side of the structure (I), A, D, B, and E arepreferably CH, and L is N or CH, with the proviso that when L is N, anysubstituents G³ are preferably monovalent, and when L is CH then anysubstituents G³ are preferably divalent.

The substituents G³ are preferably selected from the group consisting ofmonovalent moieties lower alkyl; —NR³COR⁶; —OR⁶; —SR⁶; —S(O)R⁶;—S(O)₂R⁶; —CO₂R⁶; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; —CN; optionally substitutedaryl; optionally substituted heteroaryl; optionally substitutedheteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);and bivalent bridge of structure T²═T²—T³ wherein T2 represents N or CH.T³ is preferably S, O, CR⁴ ₂, or NR³.

Most preferably, G³ is selected from the group consisting of monovalentmoieties lower alkyl; —NR³COR⁶; —CO₂R⁶; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; andbivalent bridge of structure T²═T²—T³ wherein T2 represents N or CH.Most preferably T³ is S, O, CH₂, or NR³.

Most preferably, the subscript q, which represents the number ofsubstituents G³, is 1.

Ring J is preferably a phenyl ring, and subscript q′ representing thenumber of substituents G⁴ on the phenyl ring, is preferably 0, 1, 2, or3. Subscript q′ is most preferably 1, or 2.

G⁴ moieties are preferably selected from the group consisting of—N(R⁶)₂; —NR³COR⁶; halogen; alkyl; halogen-substituted alkyl;hydroxy-substituted alkyl; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; amino-substituted alkylamino; N-loweralkylamino-substituted alkylamino; N,N-di-lower alkylamino-substitutedalkylamino; N-lower alkanoylamino-substituted alkylamino;hydroxy-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;optionally substituted heteroarylalkyl; optionally substitutedheteroaryloxy; —S(O)_(p)(optionally substituted heteroaryl); optionallysubstituted heteroarylalkyloxy; —S(O)_(p)(optionally substitutedheteroarylalkyl); as well as fused ring-forming bridges attached to andconnecting adjacent positions of the phenyl ring, said bridges havingthe structures:

 wherein each T² independently represents N, or CH; T³ represents S, orO; and binding to the phenyl ring is achieved via terminal atoms T² andT³;

 wherein each T² independently represents N, CH, or CG^(4′); with theproviso that a maximum of two bridge atoms T² may be N; and binding tothe phenyl ring is achieved via terminal atoms T²; and

 wherein each T⁵, and T⁶ independently represents O, S, or CH₂; andbinding to ring J is achieved via terminal atoms T⁵; with the provisosthat:

i) a bridge comprising T⁵ and T⁶ atoms may contain a maximum of twoheteroatoms O, S, or N; and

ii) in a bridge comprising T⁵ and T⁶ atoms, when one T⁵ group and one T⁶group are O atoms, or two T⁶ groups are O atoms, said O atoms areseparated by at least one carbon atom.

Alkyl groups which constitute all or part of a G⁴ moiety are preferablylower alkyl.

When G⁴ is an alkyl group located on ring J adjacent to the linkage—(CR⁴ ₂)_(p)—, and X is NR³ wherein R³ is an alkyl substituent, then G⁴and the alkyl substituent R³ on X may be joined to form a bridge ofstructure —(CH₂)_(p′) wherein p′ is preferably 2 or 3, with the provisothat the sum of p and p′ is 2 or 3, resulting in formation of anitrogen-containing ring of 5 or 6 members. Most preferably, the sum ofp and p′ is 2, resulting in formation of a 5-membered ring.

Most preferably, in G¹, G², G³, and G⁴, when two groups R⁶ are eachalkyl and located on the same N atom they may be linked by a bond, an O,an S, or NR³ to form a N-containing heterocycle of 5-6 ring atoms.

Preferably, when an aryl, heteroaryl, or heterocyclyl ring is optionallysubstituted, that ring may bear up to 2 substituents which areindependently selected from the group consisting of amino,mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino,lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl,hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy,halogenated lower alkylthio, —CH₂OR³, nitro, and cyano.

The method of the invention is intended to be employed for treatment ofVEGF-mediated conditions in both humans and other mammals.

The compounds may be administered orally, dermally, parenterally, byinjection, by inhalation or spray, or sublingually, rectally orvaginally in dosage unit formulations. The term ‘administered byinjection’ includes intravenous, intraarticular, intramuscular,subcutaneous and parenteral injections, as well as use of infusiontechniques. Dermal administration may include topical application ortransdermal administration. One or more compounds may be present inassociation with one or more non-toxic pharmaceutically acceptablecarriers and if desired, other active ingredients.

Compositions intended for oral use may be prepared according to anysuitable method known to the art for the manufacture of pharmaceuticalcompositions. Such compositions may contain one or more agents selectedfrom the group consisting of diluents, sweetening agents, flavoringagents, coloring agents and preserving agents in order to providepalatable preparations.

Tablets contain the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which arc suitable for themanufacture of tablets. These excipients may be, for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; and binding agents, forexample magnesium stearate, stearic acid or talc. The tablets may beuncoated or they may be coated by known techniques to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate maybe employed. These compounds may also be prepared in solid, rapidlyreleased form.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions containing the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions may alsobe used. Such excipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl-methylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example, lecithin, or condensation products of an alkylene oxidewith fatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolsuch as polyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives, for exampleethyl, or n-propyl, p-hydroxybenzoate, one or more coloring agents, oneor more flavoring agents, and one or more sweetening agents, such assucrose or saccharin.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example, sweetening, flavoring and coloringagents, may also be present.

The compounds may also be in the form of non-aqueous liquidformulations, e.g., oily suspensions which may be formulated bysuspending the active ingredients in a vegetable oil, for examplearachis oil, olive oil, sesame oil or peanut oil, or in a mineral oilsuch as liquid paraffin. The oily suspensions may contain a thickeningagent, for example beeswax, hard paraffin or cetyl alcohol. Sweeteningagents such as those set forth above, and flavoring agents may be addedto provide palatable oral preparations. These compositions may bepreserved by the addition of an anti-oxidant such as ascorbic acid.

Pharmaceutical compositions of the invention may also be in the form ofoil-in-water emulsions. The oil phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The compounds may also be administered in the form of suppositories forrectal or vaginal administration of the drug. These compositions can beprepared by mixing the drug with a suitable non-irritating excipientwhich is solid at ordinary temperatures but liquid at the rectal orvaginal temperature and will therefore melt in the rectum or vagina torelease the drug. Such materials include cocoa butter and polyethyleneglycols.

Compounds of the invention may also be administered transdermally usingmethods known to those skilled in the art (see, for example: Chien;“Transdermal Controlled Systemic Medications”; Marcel Dekker, Inc.;1987. Lipp et al. WO 94/04157 Mar. 3, 1994). For example, a solution orsuspension of a compound of Formula I in a suitable volatile solventoptionally containing penetration enhancing agents can be combined withadditional additives known to those skilled in the art, such as matrixmaterials and bacteriocides. After sterilization, the resulting mixturecan be formulated following known procedures into dosage forms. Inaddition, on treatment with emulsifying agents and water, a solution orsuspension of a compound of Formula I may be formulated into a lotion orsalve.

Suitable solvents for processing transdermal delivery systems are knownto those skilled in the art, and include lower alcohols such as ethanolor isopropyl alcohol, lower ketones such as acetone, lower carboxylicacid esters such as ethyl acetate, polar ethers such as tetrahydrofuran,lower hydrocarbons such as hexane, cyclohexane or benzene, orhalogenated hydrocarbons such as dichloromethane, chloroform,trichlorotrifluoroethane, or trichlorofluoroethane. Suitable solventsmay also include mixtures one or more materials selected from loweralcohols, lower ketones, lower carboxylic acid esters, polar ethers,lower hydrocarbons, halogenated hydrocarbons.

Suitable penetration enhancing materials for transdermal deliverysystems are known to those skilled in the art, and include, for example,monohydroxy or polyhydroxy alcohols such as ethanol, propylene glycol orbenzyl alcohol, saturated or unsaturated C₈-C₁₈ fatty alcohols such aslauryl alcohol or cetyl alcohol, saturated or unsaturated C₈-C₁₈ fattyacids such as stearic acid, saturated or unsaturated fatty esters withup to 24 carbons such as methyl, ethyl, propyl, isopropyl, n-butyl,sec-butyl isobutyl tert-butyl or monoglycerin esters of acetic acid,capronic acid, lauric acid, myristinic acid, stearic acid, or palmiticacid, or diesters of saturated or unsaturated dicarboxylic acids with atotal of up to 24 carbons such as diisopropyl adipate, diisobutyladipate, diisopropyl sebacate, diisopropyl maleate, or diisopropylfumarate. Additional penetration enhancing materials includephosphatidyl derivatives such as lecithin or cephalin, terpenes, amides,ketones, ureas and their derivatives, and ethers such as dimethylisosorbid and diethyleneglycol monoethyl ether. Suitable penetrationenhancing formulations may also include mixtures one or more materialsselected from monohydroxy or polyhydroxy alcohols, saturated orunsaturated C₈-C₁₈ fatty alcohols, saturated or unsaturated C₈-C₁₈ fattyacids, saturated or unsaturated fatty esters with up to 24 carbons,diesters of saturated or unsaturated dicarboxylic acids with a total ofup to 24 carbons, phosphatidyl derivatives, terpenes, amides, ketones,ureas and their derivatives, and ethers.

Suitable binding materials for transdermal delivery systems are known tothose skilled in the art and include polyacrylates, silicones,polyurethanes, block polymers, styrene-butadiene coploymers, and naturaland synthetic rubbers. Cellulose ethers, derivatized polyethylenes, andsilicates may also be used as matrix components. Additional additives,such as viscous resins or oils may be added to increase the viscosity ofthe matrix.

For all regimens of use disclosed herein for compounds of Formula I, thedaily oral dosage regimen will preferably be from 0.01 to 200 mg/Kg oftotal body weight. The daily dosage for administration by injection,including intravenous, intramuscular, subcutaneous and parenteralinjections, and use of infusion techniques will preferably be from 0.01to 200 mg/Kg of total body weight. The daily rectal dosage regimen willpreferably be from 0.01 to 200 mg/Kg of total body weight. The dailyvaginal dosage regimen will preferably be from 0.01 to 200 mg/Kg oftotal body weight. The daily topical dosage regimen will preferably befrom 0.1 to 200 mg administered between one to four times daily. Thetransdermal concentration will preferably be that required to maintain adaily dose of from 0.01 to 200 mg/Kg. The daily inhalation dosageregimen will preferably be from 0.01 to 10 mg/Kg of total body weight.

It will be appreciated by those skilled in the art that the particularmethod of administration will depend on a variety of factors, all ofwhich are considered routinely when administering therapeutics. It willalso be understood, however, that the specific dose level for any givenpatient will depend upon a variety of factors, including, but notlimited to the activity of the specific compound employed, the age ofthe patient, the body weight of the patient, the general health of thepatient, the gender of the patient, the diet of the patient, time ofadministration, route of administration, rate of excretion, drugcombinations, and the severity of the condition undergoing therapy. Itwill be further appreciated by one skilled in the art that the optimalcourse of treatment, i.e., the mode of treatment and the daily number ofdoses of a compound of Formula I or a pharmaceutically acceptable saltthereof given for a defined number of days, can be ascertained by thoseskilled in the art using conventional treatment tests.

GENERAL PREPARATIVE METHODS

The compounds of the invention may be prepared by use of known chemicalreactions and procedures. Nevertheless, the following generalpreparative methods are presented to aid the reader in synthesizing theKDR inhibitors, with more detailed particular examples being presentedbelow in the experimental section describing the working examples.

All variable groups of these methods are as described in the genericdescription if they are not specifically defined below. When a variablegroup or substituent with a given symbol (i.e. R³, R⁴, R⁶, G¹, G², G³,or G⁴) is used more than once in a given structure, it is to beunderstood that each of these groups or substituents may beindependently varied within the range of definitions for that symbol. Asdefined above, the compounds of the invention contain ring units each ofwhich may independently bear between 0 and 5 substituents G¹, G³, or G⁴,which are not defined as H. By contrast, it is to be noted that in thegeneral method schemes below, the G¹, G³, or G⁴ substituents are used asif their definition includes H, to show where such G¹, G³, or G⁴substituents may exist in the structures, and for ease in drawing. Nochange in the definition of G¹, G³, or G⁴ is intended by thisnon-standard usage, however. Thus, only for purposes of the generalmethod schemes below, G¹, G³, or G⁴ may be H in addition to the moietiesset forth in the definitions of G¹, G³, or G⁴. The ultimate compoundscontain 0 to 5 non-hydrogen groups G¹, G³, or G⁴.

Within these general methods the variable M is equivalent to the moiety

in which each variable group or substituent is allowed to independentlyvary within the limits defined earlier for that symbol.

Within these general methods the variable Q¹ is equivalent to the moiety

in which L is N and each other variable group or substituent is allowedto independently vary within the limits defined earlier for that symbol.

Within these general methods the variable Q² is equivalent to the moiety

in which each variable group or substituent is allowed to independentlyvary within the limits defined earlier for that symbol.

It is recognized that compounds of the invention with each claimedoptional functional group cannot be prepared with each of thebelow-listed methods. Within the scope of each method optionalsubstituents are used which are stable to the reaction conditions, orthe functional groups which may participate in the reactions are presentin protected form where necessary, and the removal of such protectivegroups is completed at appropriate stages by methods well known to thoseskilled in the art.

General Method A

The compounds of formula I-A in which X, M, and Q² are defined as above,Y is —CH₂—O—, —CH₂—S—, —CH₂—NH—, —O—, —S—, or —NH—, and R¹ and R²together with the carbons to which they are attached form a fused5-membered ring aromatic heterocycle, hal is halogen (Cl, Br, F, or Ibut preferably Cl, Br or F) are conveniently prepared according to areaction sequence as shown in Method A. Thus, a heterocycle of formulaII in which R is lower alkyl can be made by one skilled in the artaccording to the corresponding published procedures in the referencetable. In the cases of thiophene-2,3-dicarboxylic acid (table entry 1)and pyrazole-3,4-dicarboxylic acid (table entry 10), the carboxylicacids are converted to methyl or ethyl esters by treatment with thecorresponding alcohol and catalytic mineral acid (typically sulfuricacid) at reflux. The diester of formula II is treated with hydrazinehydrate to furnish intermediate III (for specific reaction conditionssee Robba, M.; Le Guen, Y. Bull. Soc. Chem. Fr. 1970 12 4317). CompoundIII is treated with a halogenating agent such as phosphorousoxychloride, phosphorous oxybromide, phosphorous pentabromide, orphosphorous pentachloride to yield dihalo intermediate IV. The dichloroor dibromo intermediates can be converted to the difluoro intermediate(when desired) by reaction with hydrogen fluoride. By using iodoreagents such as potassium iodide or tetrabutylammonium iodide insubsequent steps, the iodo intermediate is formed in the reactionmixtures without being isolated as a pure substance. Dihalo intermediateIV is treated with a nucleophile of formula V in refluxing alcohol orother suitable solvent such as tetrahydrofuran (THF), dimethoxyethane(DME), dimethylformamide (DMF), dimethylsulfoxide (DMSO), or the like tofurnish the intermediate of formula VI. Such condensations can also bedone in a melt free of solvent and can be catalyzed by acids such as HClor bases such as triethylamine or 1,8-diazobicyclo[5.4.0]undec-7-ene(DBU). The compound of formula VI is reacted with compounds of formulaVII in a suitable aprotic solvent such as DMSO, DMF or solvent freeoften with a basic catalyst such as DBU or CsCO₄, or a crown ether suchas 18-crown-6 at temperatures usually between room temperature andreflux to furnish invention compound of formula I-A. It is understoodthat the nature of the starting materials will dictate the choice ofsuitable solvents, catalyst (if used) and temperature by one skilled inthe art. Intermediates of formula V and VII are often commercial or areconveniently prepared by methods well known to those skilled in the art.For example see Martin, I., et al. Acta. Chem. Scand. 1995 49 230 forthe preparation of VII in which Y is —CH₂O— and Q² is 4-pyridylsubstituted by a 2-aminocarbonyl group (2-CONH₂).

REFERENCE TABLE FOR PREPARATION OF STARTING MATERIAL II

For diacid: Heffner, R.; Joullie, M. Synth. Commun.. 1991 21 (8 & 9)1055. The diacid can be converted to dimethyl ester by reflux inmethanol with catalytic sulfuric acid.

Erlenmeyer, H.; von Meyenburg, H. Helv. Chim. Acta.. 1937 20 204.

Commercially available

Bickel, H.; Schmid, H., Helv. Chim. Acta.. 1953 36 664.

Nicolaus, Mangoni. Gazz. Chim. Ital.. 1956 86 757.

Alder, Rickert. Chem. Ber.. 1937 70 1354.

Nicolaus, Mangoni. Gazz. Chim. Ital.. 1956 86 757.

Sice, J. J. Org. Chem.. 1954 19 70.

Tanaka, Y. Tetrahedron. 1973 29 3271.

Diacid: Tyupalo, N.; Semenyuk, T.; Kolbasina, O. Russ. J Phys. Chem.1992 66 463. The diacid can be converted to dimethyl ester by reflux inmethanol with catalytic sulfuric acid. Alternatively, the diester isprepared by reaction of dimethyl acetylenedicarboxylate withdiazomethane.

General Method B

The compounds of formula I-B in which M, X, and Q² are as defined aboveand Y is —CH₂O—, —CH₂—S—, —CH₂—NH—, —O—, —S—, or —NH— are convenientlyprepared as shown in Method B. According to a procedure described in theliterature (Tomisawa and Wang, Chem. Pharm. Bull., 21, 1973, 2607,2612), isocarbostyril VIII is reacted with PBr₅ in a melt to form1,4-dibromoisoquinoline IX. Intermediate IX is treated with anucleophile of formula V in refluxing alcohol to furnish intermediate offormula X. Such condensations can also be done in a melt free of solventand can be catalyzed by acids such as HCl or bases such as triethylamineor 1,8-diazobicyclo[5.4.0]undec-7-ene (DBU). The compound of formula Xis reacted with compounds of formula VII in a suitable aprotic solventsuch as DMSO, DMF or solvent free often with a basic catalyst such asDBU or CsCO₄ at elevated temperatures to furnish invention compound offormula I-B. This method is most useful when Y is —CH₂—S— or —S—.

General Method C

The compounds of formula I-C in which M, X, R¹, R², m and Q² are definedas above are conveniently prepared according by a reaction sequence asshown in method C. In this method m is preferably 0 and R¹ and R²together with the carbons to which they are attached form a fusedbenzene or fused 5-member ring aromatic heterocycle. Starting materialXI is either commercial or is prepared by one skilled in the art asshown in the reference table below. Starting material XI is reacted withurea or ammonia, usually at elevated temperature and pressure (in thecase of ammonia), to form imide XII. The imide is reacted with analdehyde XIII in acetic acid and piperidine at reflux to yieldintermediate XIV. Reaction of XIV with sodium borohydride in methanol orother suitable solvents according to the general procedure described byI. W. Elliott and Y. Takekoshi (J. Heterocyclic Chem. 1976 13, 597)yields intermediate XV. Treatment of XV with a suitable halogenatingagent such as POCl₃, POBr₃, PCl₅, PBr₅ or thionyl chloride yields halointermediate XVI which is reacted with nucleophile of formula V inrefluxing alcohol to furnish invention compound of formula I-C. Suchcondensations can also be done in a melt free of solvent and can becatalyzed by acids such as HCl or bases such as triethylamine or1,8-diazobicyclo[5.4.0]undec-7-ene (DBU). Alternatively, reagent V canbe condensed with intermediate XV be heating the two components withP₂O₅ in a melt to yield invention compound of structure I-C. This lastmethod is especially effective when X is an amine linker.

REFERENCE TABLE FOR PREPARATION OF STARTING MATERIALS

Commercial

Commercial

D. E. Ames and O. Ribeiro, J. Chem. Soc., Perkin Trans. 1 1975, 1390.

J. R. Carson and S. Wong, J. Med. Chem. 1973, 16, 172.

K. Yasuyuki, et al., J. Org. Chem. 1986, 51, 4150.

Schneller, et al., J. Med. Chem. 1978, 21, 990.

R. K. Robins et al., J. Org. Chem. 1963, 28, 3041.

P. Gupta, et al., J. Heterocycl. Chem. 1986, 23, 59.

R. B. Meyer, et al., J. Heterocycl. Chem. 1980 17, 159.

General Method D

The compounds of formula I-D-1 in which R¹, R², R⁶, M, X, Y, G³ and Zare defined as above and q is 0 or 1 are conveniently prepared via areaction sequence as shown in Method D. Thus, pyridine substitutedpyridazines or pyridines (I-D-1) are functionalized into substituted2-aminocarbonyl pyridines of formula (I-D-2) by the use of formamides(XVII) in the presence of hydrogen peroxide and iron salts, according toa procedure described in the literature (Minisci et al., Tetrahedron,1985, 41, 4157). This method works best when R¹ and R² togetherconstitute a fused aromatic heterocycle or fused aromatic carbocycle. Inthose cases that Z is CH and R¹ and R² do not form a fused aromatic, anisomeric side product in which Z is CCONHR⁶ can be formed and, if soformed, is removed from the desired product by chromatography.

General Method E

The compounds of formula I-E-1 and I-E-2 in which R¹, R², R⁶, M, X, Y,G³, and Z are defined as above, q is 0 or 1, and R³ is lower alkyl areconveniently prepared via a reaction sequence as shown in Method E.Thus, pyridine substituted pyridazines or pyridines (I-D-1) arefunctionalized into substituted 2-alkoxycarbonyl pyridines of formula(I-E-1) by the use of monoalkyloxalates (XVIII) in the presence of S₂O₈⁻², acid and catalytic amounts of AgNO₃, according to a proceduredescribed in the literature (Coppa, F. et al., Tetrahedron Letters,1992, 33 (21), 3057). Compounds of formula I-E-1 in which R³ is H arethen formed by hydrolysis of the ester with a base such as sodiumhydroxide in methanol/water. Compounds of formula I-E-2 in which the R⁶groups are independently defined as above, but especially includingthose compounds in which neither R⁶ is H, are conveniently prepared fromthe acid (I-E-1, R³=H) by treatment with amine XIX in the presence of acoupling agent such as DCC (dicyclohexylcarbodiimide). This method worksbest when R¹ and R² together constitute a fused aromatic heterocycle orfused aromatic carbocycle. In those cases that Z is CH and R¹ and R² donot form a fused aromatic, an isomeric side product in which Z is CCO₂R³can be formed in the first step and, if so formed, is removed from thedesired product by chromatography.

General Method F

The compounds of formula I-F in which M, Q² and X are defined as above,m is an integer of 1-5, and R¹ and R² together with the carbons to whichthey are attached form a fused 5-membered ring aromatic heterocycle canbe prepared via a reaction sequence as shown in method F. The readilyavailable heterocyclylcarboxylic acid starting material XX is reactedwith butyl lithium followed by dimethylformamide to yield the aldehydewith structure XXI. Reaction of XXI with hydrazine yields pyridazinoneXXII. Treatment of XXII with a suitable halogenating agent such asPOCl₃, POBr₃, PCl₅, PBr₅ or thionyl chloride yields a halo intermediatewhich is reacted with nucleophile of formula V in refluxing alcohol tofurnish intermediate compound of formula XXIII. Such condensations canalso be done in a melt free of solvent and can be catalyzed by acidssuch as HCl or bases such as triethylamine or1,8-diazobicyclo[5.4.0]undec-7-ene (DBU). Alternatively, reagent V canbe condensed with intermediate XXII be heating the two components withP₂O₅ in a melt to yield XXII. This last method is especially effectivewhen X is an amine linker. Formation and alkylation of the Reissertcompound XXIII with halide XXIV is done as described by the generalmethod of F. D. Popp, Heterocycles, 1980, 14, 1033 to yield theintermediate of structure XXV. Treatment of XXV with base then yieldsinvention compound I-F.

General Method G

The compounds of formula I-G in which M, Q² and X are defined as above,m is an integer of 1-4, and R¹ and R² together with the carbons to whichthey are attached form a fused 5-membered ring aromatic heterocycle canbe prepared via a reaction sequence as shown in method G. Aldehyde XXI,from method F, can be reduced with sodium borohydride to yield ahyroxyacid which is lactonized using methods well known to those skilledin the art such as with toluenesulfonyl chloride to yield lactone XXVI.Condensation of intermediate XXVI with aldehyde XIII in the presence ofa base such as sodium methoxide usually in a solvent such as methanolunder reflux yields an intermediate of structure XXVII. Reaction ofXXVII with hydrazine or preferably with hydrazine hydrate at atemperature of 100-150° C. leads to an intermediate of structure XXVIII.Conversion of intermediate XXVIII to invention compound of structure I-Gis done by methods as described in method C by using XXVIII rather thanXV.

General Method H

The compounds of formula I-H in which the R¹, R², M, X, R⁶; q and G³ aredefined as above are conveniently prepared via a reaction sequence asshown in Method H. Thus the methods described in Martin, I; Anvelt, J.;Vares, L.; Kuchn, I.; Claesson, A. Acta Chem. Scand. 1995, 49, 230-232or those of methods D or E above by substituting readily availablepyridine-4-carboxylic ester XXX for I-D-1 are used to convert XXX intoXXXI. Reduction of the ester as described by Martin, et al. above isnext done with a mild reducing agent such as NaBH₄ such that the amidesubstituent is left unchanged to yield alcohol XXXII. This alcohol isthen heated with a base such as DBU or CsCO₄ with halopyridazine VI frommethod A under anhydrous conditions to yield the invention compound withformula I-H.

General Method I

Invention compounds having formula I-I in which the R¹, R², M, X, R⁶, q,and G³ are defined as above and W is a bond or —CH₂— are convenientlyprepared via a reaction sequence as shown in Method I. This method isespecially useful when q is 1 and XXXIII is 4-chloropyridine.Alternatively, other 4-halopyridines such as 4-fluoropyridine or4-bromopyridine can be used in this process. Thus readily available4-halopyridines XXXIII are converted to intermediates of formula XXXIVby using the general procedures of methods D or E above by substitutingthe 4-halopyridine for I-D-1. Reaction of XXXIV with either potassium orsodium hydrogen sulfide yields a thiol having formula XXXV.Alternatively, the alcohol function of intermediate XXXII from method His converted to a leaving group by reaction with methanesulfonylchloride and a suitable base such as triethylamine in the cold such thatpolymeric material formation is minimized and the resultant intermediateis reacted with either potassium or sodium hydrogen sulfide to yield athiol having formula XXXVI. Either thiol have formula XXXV or formulaXXXVI is reacted with intermediate VI from method A and a suitable basesuch as diisopropylethylamine or CsCO₄ in DMF or other suitableanhydrous solvent or in the absence of solvent to yield I-D-9.

General Method J

Invention compounds such as those having formula I-J-1 or I-J-2 in whichthe R¹, R², M, X, W, and G³ are defined as above and having a sulfoxideor sulfone within the structure are conveniently prepared via a reactionsequence as shown in Method J. Reaction of compounds of this inventionthat contain a thioether group either as part of a substituent G¹, G³,or G⁴or as part of Y as shown in the representative structure I-I fromMethod I can be converted to the invention compounds with a sulfoxidemoiety such as I-J-1 by treatment with one equivalent ofm-chloroperbenzoic acid in methylene chloride or chloroform (MCPBA,Synth. Commun., 26, 10, 1913-1920, 1996) or by treatment with sodiumperiodate in methanol/water at between 0° C. and room temperature (J.Org. Chem., 58, 25, 6996-7000, 1993). The expected side productsconsisting of mixtures of various N oxides and the sulfone I-J-2 can beremoved by chromatography. The sulfone I-J-2 is obtained by the use ofan additional equivalent of MCPBA or preferably by use of potassiumpermanganate in acetic acid/water (Eur. J. Med. Chem. Ther., 21, 1, 5-8,1986) or by use of hydrogen peroxide in acetic acid (Chem. Heterocycl.Compd., 15, 1085-1088, 1979). In those cases that unwanted N oxidesbecome a significant product, they can be converted back to the desiredsulfoxides or sulfones by hydrogenation in ethanol/acetic acid withpalladium on carbon catalysts (Yaklugaku Zasshi, 69, 545-548, 1949,Chem. Abstr. 1950, 4474).

General Method K

Invention compounds having formula I-K in which the R¹, R², M, X, and Q¹are defined as above are conveniently prepared via a reaction sequenceas shown in Method K. One skilled in the art prepares starting materialsof structure XXXVII by methods known in the literature. For exampleXXXVII wherein R¹ and R² together with the carbons to which they areattached form a 2,3-substituted thiophene, furan, pyrrole,cyclopentadienyl, oxazole or thiazole are prepared using the generalchemistry given in J. Org. Chem., 1981, 46, 211 and hydrolizing theinitially formed tert-butyl ester with trifluoroacetic acid. Thepyrazole starting material can be prepared by reacting2-oxo-3-pentyn-1,5-dioic acid (J. Chem. Phys. 1974, 60, 1597) withdiazomethane. The starting material wherein R¹ and R² together with thecarbons to which they are attached form a phenyl are prepared by themethods of Cymerman-Craig et al., Aust. J. Chem. 1956, 9, 222, 225.Compounds of formula XXXVII in which R¹ and R² are lower alkyl areconveniently prepared according to procedures given in patent CH 482415(Chem. Abstr. 120261u, 1970). The crude diacid of formula XXXVII issubsequently treated with hydrazine to furnish pyridazinone XXXVIII (forspecific reaction conditions see Vaughn, W. R.; Baird, S. L. J. Am.Chem. Soc. 1946 68 1314). Pyridazinone XXXVIII is treated with achlorinating agent such as phosphorous oxychloride to yield anintermediate dichloro species which undergoes hydrolysis upon aqueousworkup to furnish chloropyridazine XXXIX. Chloro acid XXXIX is treatedwith a nucleophile of formula V in the presence of a base such as sodiumhydride in a solvent such as DMF or in the absence of a solvent. Theresultant acid XXXX is reduced with a reducing agent such as BH₃.THFaccording to the procedure of Tilley, J. W.; Coffen D. L. Schaer, B. H.;Lind, J. J. Org. Chem. 1987 52 2469. Product alcohol XXXXI is reactedwith a base and optionally substituted 4-halo-pridyl, optionallysubstituted 4-halo-pyrimidyl or optionally substituted 4-halo-pyridazyl(XXXXII) to furnish invention compound of formula I-K (for specificreaction conditions see Barlow, J. J.; Block, M. H.; Hudson, J. A.;Leach, A.; Longridge, J. L.; Main, B. g.; Nicholson, S. J. Org. Chem.1992 57 5158).

General Method L

Invention compounds having formula I-L in which the R¹, R², M, X, and Q¹are defined as above are conveniently prepared via a reaction sequenceas shown in Method L. Thus alcohol of formula XXXXI from method K isreacted with methanesulfonyl chloride in the presence of a suitable basefollowed by potassium or sodium hydrogen sulfide to yield thiol XXXXIII.The thiol is then reacted with 4-halopyridine XXXXII from method K inthe presence of a suitable base such as triethylamine to yield inventioncompound I-K. Alternatively, XXXXI is converted to halo intermediate offormula XXXXIV by methods well known to those skilled in the art and thehalide is reacted with thiol XXXXV to yield I-K. Intermediate XXXXIV canalso be converted to intermediate XXXXIII by treatment with KHS or NaHS.Reagents XXXXV are either commercially available such as4-mercaptopyridine or can be prepared by one skilled in the art such asby method I above.

EXPERIMENTAL EXAMPLE 1 Preparation of1-(4-chlorophenylamino)-4-(4-pyridylthio)isoquinoline

Step 1: Preparation of Intermediate A

A mixture of 2.90 g, 19.07 mMol of isocarbostyril and 14.40 g, 33.68mMol of phosphorus pentabromide were allowed to melt together at 140° C.The melt turned into a red liquid and after about 10 minutes thereaction mixture solidified and was cooled. The reaction mixture wascrushed up and dumped into ice water. The resulting solid was filteredand air-dried. wt. 5.50 g, 96% yield, mp.=94-96°. R_(f)=0.66 in 40%ethyl acetate in hexanes.

Step 2

A mixture of 1.00 g, 3.49 mMol of 1,4-dibromoisoquinoline (IntermediateA) from step 1 and 4-chloroaniline were melted together at 140°. Thereaction mixture turned into a deep red liquid and after about 10minutes the reaction mixture solidified and was done. The reactionmixture was broken up and triturated with a 50/50 methanol/THF mixturethen filtered and air dried without further purification. wt. 0.75 g,64.4%, mp.=260-263°. R_(f)=0.58 in 40% ethyl acetate in hexanes.

Step 3

A mixture of 0.05 g, 0.1498 mMol of1-(4-chloroaniline)-4-bromoisoquiniline and 0.02 g, 0.18 mMol of4-mercaptopyridine were combined and melted together at 140° for about10 minutes. The resulting reaction mixture was purified on a 1000 micronprep plate using 5% methanol in hexanes as the solvent. wt. 0.0103 g,19% yield, mp. 192-195°. R_(f)=0.50 in 40% ethyl acetate in hexanes.

EXAMPLE 2 Preparation of1-(indan-5-ylamino)-4-(4-pyridylthio)isoquinoline

The procedure used for the preparation of Example 1 was used to preparethe title compound by substituting 5-aminoindane for 4-chloroaniline instep 2. Mp. 100-103°, TLC R_(f) 0.40 (40% ethyl acetate in hexanes).

EXAMPLE 3 Preparation of1-(benzothiazol-6-ylamino)-4-(4-pyridylthio)isoquinoline

The procedure used for the preparation of Example 1 was used to preparethe title compound by substituting 6-aminobenzothiazole for4-chloroaniline in step 2.

TLC R_(f) 0.36 (5%methanol/methylene chloride); MS=387

EXAMPLE 4 Preparation of1-(4-chlorophenylamino)-4-(4-pyridylmethyl)isoquinoline

Step 1

A mixture of homophthalimide (770 mg, 4.78 mmol),4-pyridinecarboxaldehyde (0.469 mL, 4.78 mmol) and piperidine (0.5 mL)in acetic acid (25 mL) was heated at reflux for 1 h. The resultantsolution was cooled to room temperature. The solid product was removedby filtration, washed by water (4×10 mL) and dried under vacuum toafford 920 mg (3.67 mmol, 77% yield ) of a mixture of Z and E isomers ofthe above compound. ¹H-NMR (DMSO-d₆) complex proton signals shown inaromatic region due to existence of both E and Z isomers. MS ES 251(M+H)⁺, 252 (M+2H)⁺.

Step 2

To a suspension of starting material (1.70 g, 6.8 mmol) in methanol (250mL) at 0° C. was added slowly sodium borohydride (3.0 g, 79 mmol). Themixture was allowed warmed to rt and continued stirring for 1 hr. Thereaction was quenched with water (10 mL) and stirred for 10 minutes. Theresulting mixture was concentrated to remove solvent. To the residue wasadded water with ice (100 mL), and adjusted the pH=2 with 2 N HClsolution. Stirred for 10 minutes, added 2 N NaOH until pH of thesolution was about 11. The resulting solution was extracted by CH₂Cl₂(4×100 mL). The combined organic layers were collected, dried over MgSO₄and concentrated. The residue was purified by column chromatography(1:10 v/v methanol-dichloromethane) to afford 400 mg of the titlecompound as a solid (1.70 mmol, yield 25%). ¹H-NMR (MeOH-d₄) 8.33 to8.39 (m, 4H), 7.50 to 7.68 (m, 3H), 7.30-7.31 (m, 2H), 7.14 (s, 1H),4.15 (s, 2H); MS ES 237 (M+H)+, 238 (M+2H); TLC (1:10 v/vmethanol-dichloromethane) R_(f)=0.40.

Step 3

A mixture of 4-chloroaniline (178 mg, 1.40 mmol), phosphorus pentoxide(396 mg, 1.40 mmol) and triethylamine hydrochloride (193 mg, 1.40 mmol)was heated and stirred under argon at 200° C. for 1.5 h or until ahomogenous melt has formed. To the melt was added starting material (82mg, 0.35 mmol). The reaction mixture was stirred at 200° C. for 2 h. Theresultant solid black mass was cooled to 100° C. Methanol (5 mL) andwater (10 mL) were added and the reaction mixture was sonicated untilthe black mass had become soluble. Dichloromethane (40 mL) was added andconcentrated ammonia (2 mL) was added to adjust the mixture to pH=10.The organic layer was separated, and the aqueous layer was extractedwith dichloromethane (3×20 mL). The combined organic layers were driedover MgSO₄, filtered, and concentrated. Purification by preparative TLCplate (1:10 v/v methanol-dichloromethane) yielded 26 mg (0.08 mmol, 22%yield) of the title compound as a yellow solid. ¹H-NMR (MeOH-d₄) 8.37(d, J=7.8 Hz, 3H), 7.86 (s, 1H), 7.55 to 7.77 (m, 5H), 7.27 to 7.33 (m,4H), 4.31 (s, 2H); MS ES 346 (M+H)⁺; TLC (1:10 v/vmethanol-dichloromethane) R_(f)=0.45.

EXAMPLE 5 Preparation of1-(benzothiazol-6-ylamino)-4-(4-pyridylmethyl)-isoquinoline

The procedure used for the preparation of Example 4 was used to preparethe title compound by substituting 6-aminobenzothiazole for4-chloroaniline in step 3. ¹H-NMR (MeOH-d₄) 9.08 (s, 1H), 8.37 to 8.59(m, 4H), 7.79 to 8.01 (m, 2H), 7.60 to 7.78 (m, 4H), 7.30 (d, 2H), 4.34(s, 2H); MS ES 369 (M+H)⁺; TLC (1:4 v/v hexane-ethyl acetate)R_(f)=0.20.

EXAMPLE 6 Preparation of1-(indan-5-ylamino)-4-(4-pyridylmethyl)-isoquinoline

The procedure used for the preparation of Example 4 was used to preparethe title compound by substituting 5-aminoindane for 4-chloroaniline instep 3. ¹H-NMR (MeOH-d₄) 8.35 (m, 3H), 7.46 to 7.77 (m, 5H), 7.15 to7.27 (m, 4H), 4.26 (s, 2H), 2.87 to 2.90(m, 4H), 2.05 to 2.10 (m, 2H);MS ES 352 (M+H)⁺; TLC (1:4 v/v hexane-ethyl acetate) R_(f)=0.25.

EXAMPLE 7 Preparation of1-(3-fluoro-4-methylphenylamino)-4-(4-pyridylmethyl)-isoquinoline

The procedure used for the preparation of Example 4 was used to preparethe title compound by substituting 3-fluoro-4-methylaniline for4-chloroaniline in step 3. ¹H-NMR (MeOH-d₄) 8.34 (d, 3H), 7.87 (s, 1H),7.54 to 7.69 (m, 4H), 7.10 to 7.31 (m, 4H), 2.22 (s, 3H); MS ES 344(M+2H)⁺; TLC (1:4 v/v hexane-ethyl acetate) R_(f)=0.20.

EXAMPLE 8 Preparation of4-(4-chlorophenylamino)-7-(4-pyridylmethoxy)thieno-[2,3-d]pyridazine

Step 1

A dry, 2 L, 3-necked, round-bottomed flask was equipped with amechanical stirrer and addition funnel. To the flask was added2-thiophenecarboxylic acid (25 g, 195 mmol) in anhydrous THF (500 mL)under argon. The mixture was cooled to −78° C. with a dryice-isopropanol bath and allowed to stir for 30 min. n-Butyllithium inHexanes (2.5 M, 172 mL) was added dropwise over 30 min. The reaction waskept at −78° C. for an additional hour with stirring then placed underan atmosphere of dry carbon dioxide. With addition of the carbon dioxidethe reaction became thick. The reaction remained at −78° C. for anadditional hour before warming to −10° C. The reaction was quenched with2 N HCl (213 mL) and allowed to reach rt. The layers were separated andthe aqueous layer was extracted with EtOAc (3×200 mL). The organiclayers were combined, dried (Na₂SO₄) and concentrated by rotaryevaporation. The brown solid was crystallized from hot isopropanol anddried overnight under vacuum. Desired thiophene-2,3-dicarboxylic acidwas obtained (27.3 g, 159 mmol; 82% yield); ¹H NMR (DMSO-d₆) 7.69 (d,J=1.5, 1), 7.38 (d, J=4.8, 1); ES MS (M+H)⁺=173; TLC(Chloroform-MeOH-water, 6:4:1); R_(f)=0.74.

Step 1A

Alternatively, 3-thiophenecarboxylic acid rather than2-thiophenecarboxylic acid has been used in step 1 to yield the sameproduct.

Step 2

A 1 L, round-bottomed flask was equipped with a stir bar and refluxcondenser. To the flask was added the product of step 1 (62 g, 360 mmol)in MeOH (500 mL) with a catalytic amount of H₂SO₄ (˜5 mL). The reactionwas heated to reflux and stirred for 24 h. The reaction was cooled to rtand concentrated rotary evaporation. The brown mixture was purified bysilica gel chromatography (Hexane-EtOAc 80:20 gradient to 60:40).Desired dimethyl thiophene-2,3-dicarboxylate was obtained (21.2 g, 106mmol; 31% yield); ¹H NMR (DMSO-d₆) 7.93 (d, J=4.8, 1), 7.35 (d, J=4.8,1), 3.8 (d, J=1, 6); ES MS (M+H)⁺=201; TLC (Hexane-EtOAc, 70:30);R_(f)=0.48.

Step 3

A 250mL, round-bottomed flask was equipped with a stir bar and refluxcondenser. To the flask was added the product of step 2 (16 g, 80 mmol),hydrazine hydrate (6.6 mL, 213 mmol), and EtOH (77 mL) and refluxed for2.5 h. The reaction was cooled to rt and concentrated by rotaryevaporation. Water (50 mL) was added and the filtrate was separated fromthe insoluble solids. The aqueous layer was concentrated by rotaryevaporation to give a pale yellow solid. The solid was dried in a vacuumoven overnight at 50° C. Desired thieno[2,3-d]pyridazin-4,7-dione wasobtained (12 g, 71 mmol; 89% yield); ¹H NMR (DMSO-d₆) 7.85 (d, J=5.1,1), 7.42 (d, J=5.1, 1); ES MS (M+H)⁺=169; TLC (dichloromethane-MeOH,60:40); R_(f)=0.58.

Step 4: Preparation of Intermediate B

A 250 mL, round-bottomed flask was equipped with a stir bar and refluxcondenser. To the flask was added the product of step 3 (2.5 g, 14.8mmol), phosphorus oxychloride (45 mL, 481 mmol), and pyridine (4.57 mL,55 mmol) and refluxed for 2.5 h. The reaction was cooled to rt andpoured over ice. The mixture was separated and the aqueous layer wasextracted with chloroform (4×75 mL). The organic layers were combined,dried (Na₂SO₄) and concentrated by rotary evaporation to give a darkyellow solid. Desired 4,7-dichlorothieno[2,3-d]pyridazine (IntermediateB; 1.5 g, 7.3 mmol; 49% yield); mp=260-263° C.; ¹H NMR (DMSO-d₆) 8.55(d, J=5.7, 1), 7.80 (d, J=5.7, 1); ES MS (M+H)⁺=206; TLC (hexane-EtOAc,70:30); R_(f)=0.56. See also Robba, M.; Bull. Soc. Chim. Fr.; 1967,4220-4235.

Step 5

A 250 mL, round-bottomed flask was equipped with a stir bar and refluxcondenser. To the flask was added the product of step 4 (7.65 g, 37.3mmol), 4-chloroaniline (4.76, 37.3 mmol) in EtOH (75 mL). The mixturewas refluxed for 3 h. An orange solid precipitated from the reactionafter 3 h. The reaction was cooled to rt and the solid was collected byfiltration and washed with hexane. The desired7-chloro-4-(4-chlorophenylamino)thieno[2,3-d]pyridazine was obtained(6.5 g, 21.9 mmol; 60% yield); mp=139-142° C.; ES MS (M+H)⁺=297; TLC(Hexane-EtOAc, 60:40); R_(f)=0.48.

Step 6

A 150 mL, round-bottomed flask was equipped with a stir bar and refluxcondenser. To the flask was added the product of step 5 (0.33 g, 1.1mmol), 4-pyridylcarbinol (1.2 g, 11.2 mmol) in DBU (2.5 mL, 16.7 mmol)and the mixture was heated to 125° C. for 24 hours. EtOAc (10 mL) wasadded to the reaction while hot and then the reaction was poured intowater (10 mL). The layers were separated and the aqueous layer wasextracted with EtOAc (3×10 mL). The organic layers were combined, dried(MgSO₄) and concentrated by rotary evaporation. The resulting mixturewas purified by silica gel chromatography(dichloromethane-methanol-acetone, 90:5:5) to give a pale yellow solid.The desired title compound was obtained (0.03 g, 0.08 mmol; 7.3% yield);mp=203-205° C. dec; ES MS (M+H)⁺=369; TLC(dichloromethane-methanol-acetone, 95:2.5:2.5); R_(f)=0.56.

EXAMPLE 9 Preparation of4-(4-chlorophenylamino)-7-(4-pyridylmethoxy)furo[2,3-d]pyridazine

Step 1

n-Butyllithium (2.5M in hexanes, 196 mL, 491 mmol) was introduced into adry 3 L 3-necked flask fitted with an addition funnel, argon inlet, andmechanical stirrer. The mixture was diluted with dry THF (500 mL), andcooled to −78° C. 3-furoic acid (25 g, 223 mmol) was added as solutionin THF (500 mL) dropwise. The mixture was stirred for 1.5 h, at whichpoint dry carbon dioxide was bubbled through the reaction mixture for 1h. After warming gradually to −10° C., the resultant thick white slurrywas treated with aqueous HCl (2 N, 446 mL). The two layers wereseparated, and the aqueous layer was extracted with EtOAc (3×300 mL).The combined organics were dried (Na₂SO₄), filtered, and concentrated toafford crude furan-2,3-dicarboxylic acid as an orange solid (44 g) whichwas used without further purification. ¹H NMR (300 MHz, d₆-acetone)δ7.06 (d, J=1.7, 1), 7.97 (d, J=1.7, 1), 10.7 (bs, 2H);TLC(CHCl₃/MeOH/H₂O 6:4:1) R_(f)=0.56.

Step 2

A dry 500 mL round bottomed flask was equipped with a stir bar and anargon inlet. The flask was charged with the crude diacid prepared inStep 1 (44 g) dissolved in MeOH (250 mL). To the reaction mixture wasadded chlorotrimethylsilane (80 mL, 630 mmol) portionwise. Afterstirring at room temperature for 15.5 h, the solution was concentratedto an oil and silica (5 g) was added. The mixture was suspended in MeOH(100 mL), and the volatiles were removed. Suspension in MeOH (100 mL)and the removal of the volatiles was repeated an additional two times.The residue was applied directly to the top of a flash chromatographycolumn and was eluted hexanes/EtOAc 60:40 to yield dimethylfuran-2,3-dicarboxylate as an orange oil (38 g, 93% for Step 1 and Step2 combined). ¹H NMR (300 MHz, CDCl₃) δ 3.81 (s, 3), 3.86 (s, 3), 6.71(d, J=2.8, 1), 7.46 (d, J=2.8, 1); TLC (hexanes/EtOAc 60:40) R_(f)=0.46.

Step 3

A 500 mL round bottomed flask fitted with an argon inlet, a refluxcondenser, and a stir bar was charged with dimethylfuran-2,3-dicarboxylate (44 g, 236 mmol) dissolved in EtOH (250 mL).Hydrazine hydrate (55% N₂H₄, 40 mL, 3.0 mmol) was added to the solution,and the reaction mixture was warmed to reflux. A yellow solid slowlyprecipitated over the course of 5.5 h, at which point the mixture wascooled to room temperature. The volatiles were removed under reducedpressure to furnish a yellow paste which was suspended in water andfiltered. The yellow solid was washed with water and transferred to a500 mL round bottomed flask fitted with an argon inlet, a refluxcondenser, and a stir bar. The solid was suspended in aqueous HCl (2N,200 mL), and the mixture was warmed to reflux. After heating for 4 h,the orange slurry was cooled to room temperature and filtered. The solidwas washed thoroughly with water and dried under vacuum to yield4,7-dioxo[2,3-d]furopyridazine as an orange solid (21.5 g, 60%). ¹H NMR(300 MHz, d₆-DMSO) δ 7.00 (d, J=2.1, 1), 8.19 (d, J=2.1, 1H), 11.7 (bs,2H).

Step 4: Preparation of Intermediate C

A 1 L round bottomed flask was fitted with a reflux condenser, a stirbar, and an argon inlet. The furan from Step 3 (15.5 g, 102 mmol) wasadded to a mixture of phosphorous oxychloride (300 mL) and pyridine (30mL), and the resultant orange suspension was warmed to reflux. Afterheating the reaction mixture for 4 h, the volatiles were removed byrotary evaporation. The residue was poured onto ice, and the aqueousmixture was extracted with CHCl₃ (4×250 mL). The combined organics werewashed with brine, dried (MgSO₄) and concentrated to afford4,7-dichloro[2,3-d]furopyridazine (Intermediate C, 11.3 g, 59%) as anorange-red solid which was used without further purification. TLC(hexanes/EtOAc) R_(f)=0.352; ¹H NMR (300 MHz, d₆-DMSO) δ 7.40 (d, J=2.0,1), 8.63 (d, J=2.0, 1).

Step 5

A 100 mL round bottomed flask fitted with a stir bar, an argon inlet,and a reflux condenser was charged with the product of Step 4 (1.50 g,7.98 mmol) dissolved in ethanol (40 mL). Chloroaniline was added to thismixture (1.02 g, 7.98 mmol), and the resultant suspension was warmed toreflux. After heating for 4 h, the mixture was concentrated by rotaryevaporation. The crude orange solid was applied to the top of a flashcolumn and eluted with CH₂Cl₂/MeOH 97:3 to afford a mixture of4-chloro-7-[N-(4-chlorophenyl)amino][2,3-d]furopyridazine and7-chloro-4-[N-(4-chlorophenyl)amino][2,3-d]furopyridazine as a yellowpowder (1.2 g, 55%). TLC (CH₂Cl₂/MeOH 97:3); R_(f)=0.7; ¹H NMR (300 MHz,d₆-DMSO) δ major isomer (A) 7.40 (d, J=8.9, 2), 7.45 (d, J=2.0, 1), 7.87(d, J=9.2, 2), 8.34 (d, J=2.0, 1) 9.62 (s, 1); minor isomer (B) 7.28 (d,J=2.0, 1), 7.40 (d, J=8.9, 2), 7.87 (d, J=9.2, 2), 8.48 (d, J=2.1, 1),9.88 (s, 1).

Step 6

A 25 mL round bottomed flask was fitted with an argon inlet, a stir bar,and a reflux condenser. The product of step 5 (400 mg, 1.4 mmol) wascombined with 4-pyridylcarbinol (782 mg, 7.17 mmol) and1,8-diazabicyclo[5.4.0]undec-7-ene (2.5 mL 16.7 mmol), and the slurrywas warmed to 125° C. After stirring for 24, the reaction was cooled,applied directly to the top of a flash column, and eluted withCH₂Cl₂/MeOH 95:5. The resultant yellow oil was rechromatographed underthe same conditions to yield the title compound as part of a mixture ofthree components. HPLC separation (C₁₈ column CH₃CN/H₂O 10:90 gradientto 100:0) furnished the title compound as an off white solid (13.7 mg,3%). TLC (CH₂Cl₂/MeOH 95:5)=0.19; MP 198° C.; ¹H NMR (300 MHz, CDCl₃) δ5.60 (s, 2), 6.6 (d, J=2.1, 1), 7.18-7.20 (m, 2), 7.35-7.43 (m, 6), 7.66(d, J=2.1, 1) 8.54 (d, J=5.6, 2).

Steps 5A and 6A

Alternatively 4,7-dibromo[2,3-d]furopyridazine (Intermediate G below) isused to prepared the title compound by following step 5 but substitutingthe dibromo intermediate for the dichloro intermediate. Step 6A isconducted by melting the two components together in the presence ofCsCO₄ rather than 1,8-diazabicyclo[5.4.0]undec-7-ene. The crude productis purified as above.

Intermediates D to G Preparation of Other Bicyclic4,5-fused-3,6-dihalopyridazines

The general procedures of example 9, steps 2 to 4 are used bysubstituting the appropriate heterocycledicarboxylic acid forfuran-2,3-dicarboxylic acid to yield the substituted dichloropyridazinesD to G found in the below table. The dibromofuropyridazine G wasprepared using steps 2-3 from example 9 and then conducting step 4′ asfollows: to 0.50 g (3.287 mmol) of the product of step 3 was added 2.83g (6.57 mmol) of phosphorus pentabromide. This was heated to 125° C. Atabout 115° C. the reaction mixture melted and then re-solidified beforeit reached 125° C. The reaction mixture was cooled and the solid residuewas crushed up and dumped into ice water. The resulting solid was thenfiltered and vacuum dried. wt.=0.75 g (82% yield). In several cases thedichloropyridazines are known materials, as indicated by the givenreference. All of these dihaloheterocycles can be used to prepare theclaimed invention compounds.

TABLE D

Was prepared according to methods of: Robba, M.; Bull. Soc. Chim. Fr.;263, 1966, 1385-1387 1H NMR (DMSO-d6) 9.94(s, 1); ES MS (M + M)+ = 207 E

Was prepared: 1H NMR (DMSO-d6) 8.85(s, 1); ES MS(M + H)+ = 189 F

Can be prepared using the methods of: Robba, M., et.al; Bull. Soc. Chim.Fr.; 1967, 4220-4235 G

TLC R_(f) 0.76 (5% MEOH/methylene chloride)

Intermediate H Preparation of (2-methylaminocarbonyl-4-pyridyl)methanol

Step 1

A stirred solution of ethyl isonicotinate (250 mL, 1.64 mole) andconcentrated sulfuric acid (92 mL, 1.64 mole) in N-methylformamide (2.0L) was cooled to 6° C. with an ice bath. Iron (II) sulfate heptahydrate(22.8 g, 0.0812 mole, milled with a mortar and pestle) was added,followed by the dropwise addition of 30% aqueous hydrogen peroxide (56mL, 0.492 mole). The additions of iron (II) sulfate and hydrogenperoxide were repeated four additional times, while the reactiontemperature was kept below 22° C. After the reaction mixture was stirredfor thirty minutes, sodium citrate solution (2 L, 1 M) was added (pH ofthe resulting mixture was about 5). The mixture was extracted withdichloromethane (1L, 2×500 mL). The combined organic extracts werewashed with water (2×500 mL), 5% aqueous sodium bicarbonate (3×100 mL),and brine (500 mL). The resulting organic solution was then dried oversodium sulfate, filtered and concentrated in vacuo to afford a solid.The crude solid was triturated with hexanes, filtered, washed withhexanes and dried under vacuum to give 270.35 g (79.2%) of pastel yellowsolid. ¹H NMR (DMSO-d₆, 300 MHz): δ 8.9 (d, 1H), 8.3 (m, 1H), 8.0 (dd,1H), 4.4 (q, 2H), 2.8 (d, 3H), 1.3 (t, 3H).

Step 2

To a mechanically stirred slurry of the product of step 1 (51.60 g,0.248 mole) in EtOH (1.3 L) was added sodium borohydride (18.7 g, 0.495mole). The reaction mixture was stirred at rt for 18 hr. The resultingsolution was quenched carefully with saturated aqueous ammoniumhydrochloride (2 L). Gas evolution was observed during quenching. Theresulting mixture was basified with conc. ammonium hydroxide solution(200 ml) to pH=9. It was then extracted with EtOAc (8×400 mL). Thecombined organic layers were dried (MgSO₄), filtered, and concentratedin vacuo to give Intermediate H as a clear light yellow oil (36.6 g, 89%yield). ¹H NMR (DMSO-d₆, 300 MHz): δ 8.74 (q, 1H), 8.53 (dd, 1H), 7.99(m, 1H), 7.48 (m, 1H), 5.53 (t, 1H), 4.60 (d, 2H), 2.81 (d, 3H); MS m/z167 [M+H]⁺.

Intermediates I to N General Method for Preparation of[2-(N-Substituted)aminocarbonyl-4-pyridyl]methanol Intermediates

To a 0° C. solution of the amine 2 (3 equiv) in benzene is addedtrimethyl aluminum (3 equiv). Gas evolution is observed and the reactionis then allowed to warm to rt and stir for 1 h. (Lipton, M. F. et al.Org. Synth. Coll. Vol. 6, 1988, 492 or Levin, J. I. et al. Synth. Comm.,1982, 12, 989). The known carbinol 1 (1 equiv, Hadri, A. E.; Leclerc, G.Heterocyclic Chem, 1993, 30, 631) is added to the aluminum reagent andthe mixture is heated to reflux for 1 h. The reaction is quenched withwater and concentrated. The crude product is usually purified by silicagel column chromatography (20/1 EtOAc/MeOH) to afford title compound 3.The final products are generally confirmed by LC/MS and NMRspectroscopy.

Example Amine 2 Used Characterization of Compound 3 I

(M + H)⁺ 223 R_(f) = 0.17 (100% EtOAc) J

(M + H)⁺ 181 R_(f) = 0.2 (9:1 EtOAc/MeOH) K

(M + H)⁺ 224 R_(f) = 0.14 (1:1 EtOAc/CH₂Cl₂) L

(M + H)⁺ 193 R_(f) = (0.58 100% EtOAc) M

(M + H)⁺ 311 R_(f) = 0.34 (3/2 EtOAc/Hex) N

(M + H)⁺ 181 R_(f) = 0.46 (100% EtOAc) * CH₂Cl₂ is used as the solventrather than benzene.

EXAMPLE 10 Preparation of4-(4-chlorophenylamino)-7-(2-aminocarbonyl-4-pyridylmethoxy)thieno-[2,3-d]pyridazine

A 25 mL, 3-necked, round-bottomed flask was equipped with a stir bar andthermometer. To the flask was added the product of Example 8 (0.475 g,1.29 mmol), iron sulfate heptahydrate (0.179 g, 0.64 mmol), formamide(11.15 mL, 281 mmol) and conc. H₂SO₄ (0.14 mL). The mixture was stirredfor 30 min at rt at which time H₂O₂ (0.2 mL, 6.44 mmol) was added dropwise to the mixture. The reaction stirred at room temperature for anadditional hour and then heated to 55° C. over 30 min. The reaction waskept at this temperature for 3 h and then cooled to rt. An aqueoussolution of sodium citrate (0.27M, 1 mL) was added to the reaction andsubsequently the layers were separated and the aqueous layer wasextracted with EtOAc (4×5 mL). The organic layers were combined, dried(MgSO₄) and concentrated by rotary evaporation. The resulting solid wastaken up in hot acetone and separated from any remaining solids byfiltration. The filtrate was then concentrated by rotary evaporation andthe resulting residue was taken up in hot MeOH and the white solid wascollected by filtration. Desired compound (0.014 g, 0.034 mmol; 2.7%yield); mp=233° C.; ES MS (M+H)⁺=412; TLC(dichloromethane-methanol-acetone, 95:2.5:2.5); R_(f)=0.20.

EXAMPLE 11 Preparation of4-(4-chlorophenylamino)-7-(2-methylaminocarbonyl-4-pyridylmethoxy)thieno-[2,3-d]pyridazine

The procedure used for the preparation of Example 10 was used to preparethe title compound by substituting methylformamide for formamide: 1H NMR(DMSO-d6) 8.80 (d, 1), 8.62 (d,1), 8.31 (d, 1), 8.09 (d, 2), 7.86 (d,2), 7.65 (d, 1), 7.35 (d, 2), 5.74 (s, 2), 2.84 (d, 3); ES MS (M+H)⁺=426(ES); R_(f) (95/2.5/2.5 DCM/MeOH/Acetone)=0.469.

EXAMPLE 12 Preparation of1-(4-chlorophenylamino)-4-(2-aminocarbonyl-4-pyridylmethyl)isoquinoline

The procedure used for the preparation of Example 10 was used to preparethe title compound by substituting the product of example 4 for theproduct of example 8. The crude product was purified by preparative TLCplate (1:4 v/v hexane-ethyl acetate, 19% yield) of the title compound asa yellow solid. ¹H-NMR (MeOH-d₄) 8.42 (d, 1H), 8.34 (d, 1H), 7.94 (s,1H), 7.88 (s, 1H), 7.55 to 7.76 (m, 5H), 7.26 to 7.36 (m, 3H), 4.34 (s,2H); MS ES 389 (M+H)⁺; TLC (1:4 v/v hexane-ethyl acetate) R_(f)=0.44.

EXAMPLE 13 Preparation of1-(4-chlorophenylamino)-4-(2-methylaminocarbonyl-4-pyridylmethyl)isoquinoline

The procedure used for the preparation of Example 11 was used to preparethe title compound by substituting the product of example 4 for theproduct of Example 8. The crude product was purified by columnchromatography (2:3 v/v hexane-ethyl acetate, 20% yield) of the titlecompound as a yellow solid. ¹H-NMR (MeOH-d₄) 8.42 (d, 1H), 8.33 (d, 1H),7.88 (d, 2H), 7.55 to 7.77 (m, 5H), 7.28 to 7.36 (m, 3H), 4.34 (s, 2H),2.89 (s, 3H); MS ES 403 (M+H)⁺; TLC (2:3 v/v hexane-ethyl acetate)R_(f)=0.30.

EXAMPLES 14 AND 15 Preparation of4-(4-chlorophenylamino)-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazineand4-(4-chlorophenylamino)-2-methylaminocarbonyl-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine

To a suspension of the final product from Example 9 (19.20 g, 54.4 mmol)in N-methylformamide (200 mL) and distilled water (20 mL) at roomtemperature was added concentrated H₂SO₄ (2.9 mL, 54.4 mmol) dropwise.The mixture was stirred until it became a clear solution. To thissolution was added FeSO₄.7H₂O (1.51 g, 5.43 mmol) in one portion,followed by the addition of hydroxylamine-O-sulfonic acid (HOSA, 1.84 g,16.3 mmol). The additions of FeSO₄.7H₂O and HOSA were repeated in 10min. intervals for 11 times. HPLC assay showed the consumption of moststarting material. The reaction mixture was cooled with an ice bath. Asolution of sodium citrate (600 mL, 1M, 600 mmol) was added undervigorous stirring. The resulting suspension was stirred vigorously foradditional 10 min. The solid was collected by filtration, washed withwater (3×100 mL), and dried under vacuum at 50° C. for 16 hours. Thecrude product (21 g) was purified by filtering through a silica gel padeluting with 5% CH₃OH/CH₂Cl₂. The resulting 3.7 g product wasrecrystallized in CH₃CN (125 mL, boiled for 1.5 hours). The solid wascollected by filtration, washed with CH₃CN (2×15 mL), and dried undervacuum at 50° C. for 16 hours. The final product(4-(4-chlorophenylamino)-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine)is a light yellow solid (3.38 g, 15.2%). mp=223-224° C.

A major byproduct was isolated through the above silica gel padfiltration. The structure of the byproduct(4-(4-chlorophenylamino)-2-methylaminocarbonyl-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine)was characterized by ¹H NMR, 2D NMR, elemental analysis, and MS. ¹H NMR(DMSO-d₆, 300 MHz): δ 9.32 (br s, 1H), 8.93 (q, 1H), 8.79 (q, 1H), 8.63(dd, 1H), 8.12 (m, 1H), 7.91 (m, 3H), 7.70 (dd, 1H), 7.35 (m, 2H), 5.76(br s, 2H), 2.81 (d, 6H). MS m/z 467 [M+H]⁺.

EXAMPLE 14A Preparation of4-(4-chlorophenylamino)-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine—Process2

To a mixture of the Intermediate from Example 9, step 5 (10.0 g, 35.7mmol), Intermediate H (12.4 g, 74.6 mmol), and 18-crown-6 (0.42 g, 1.59mmol) in toluene (100 mL) was added KOH powder (4.4 g, 85%, 66.7 mmol)in one portion at room temperature. The reaction mixture was then heatedto 85±2° C. under vigorous stirring. The reaction mixture was stirredvigorously at this temperature overnight. After it was cooled to roomtemperature, toluene solution was decanted off and water (100 mL) wasadded to the gummy residue. The resulting mixture was stirred vigorouslyuntil it became a free flowing suspension. The solids were collected byfiltration, washed with water (2×10 mL), and dried under vacuum at 45°C. for 16 hours. The yellow/brown solids were suspended in acetonitrile(70 mL) and the suspension was stirred at reflux for 2 hours. After itwas cooled to room temperature, the solids were collected by filtration,washed with small amount of acetonitrile, and dried under vacuum at 45°C. overnight. The title product was isolated in 46% yield (6.73 g) as alight yellow solid.

EXAMPLE 16 Preparation of4-(4-chlorophenylamino)-7-(2-aminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine

The procedure used for the preparation of Example 14 was used to preparethe title compound by substituting Formamide for N-methylformamide. Thereaction was conducted with 500 mg of final product from Example 9 andproportional amounts of solvents and reagents. The crude product waspurified by HPLC on a 75×30 mm C18 column and a linear gradient elutionfrom 10 to 100% acetonitrile in water with 0.1% trifluoroacetic acid at10 ml/min. over 10 min. to yield 18 mg of the title compound as a yellowsolid: HPLC (50×4.6 mm YMC CombiScreen® C18 column, linear gradient 10to 100% acetonitrile in water with 0.1% trifluoroacetic acid at 3ml/min. over 5 min., UV detection at 254 nm) 2.35 min. peak; MS ES 396(M+H)⁺.

EXAMPLE 17 Preparation of4-(4-chlorophenylamino)-7-(benzothiazol-6-ylamino)thieno[2,3-d]pyridazine

To the dichloride from Example 8, step 4 (1.00 g, 4.90 mmol) was addedp-chloroaniline (622 mg, 4.90 mmol) and absolute ethyl alcohol (10.0mL). The mixture was refluxed at 95° C. for 2 hrs and then cooled toroom temperature. The yellow precipitate (2) that formed was filteredand washed with isopropyl alcohol, 4.0 N KOH, H₂O, and then hexane. Thefiltrate (2) was then mixed 6-aminobenzothiazole (883 mg, 5.88 mmol) in10 mL of n-butanol, and heated at 150° C. overnight. The reaction wasallowed to cool to room temperature before the solvent was removed byrotary evaporation. The residue was treated sequentially with aqueous4.0 N KOH solution and extracted with dichloromethane (50 mL), dried(MgSO₄), and the solvent evaporated. The crude product was purified byflash chromatography on silica gel using 95% dichloromethane/methanol asthe eluent. The structure of the pure title compound was confirmed byLC/MS and NMR: TLC (30% EtOAc/Hexanes) R_(f) (3)=0.20; ¹H NMR (DMSO) δ7.2 (dd, 3H), 7.38 (dd, 3H), 7.65 (d, 1H), 8.0 (d, 1H), 8.45 (d, 1H),8.8 (s, 1H); LC/MS m/z 410 rt=4.21 min.

EXAMPLE 18 Preparation of4-(indan-5-ylamino)-7-(benzothiazol-6-ylamino)thieno-[2,3-d]pyridazine

The procedure used for the preparation of Example 17 was used to preparethe title compound by substituting 5-aminoindane for 4-chloroaniline.The crude product was purified by flash chromatography on silica gelusing 30% ethyl acetate/hexane as the eluent. The structure of the puretitle compound was confirmed by LC/MS and NMR: TLC (30% EtOAc/Hexanes)R_(f) (3)=0.20; (3) ¹H NMR (DMSO) δ 2.0 (m, 2H), 2.85 (m, 4H), 7.18 (d,1H), 7.8 (d, 1H), 7.95 (d, 1H), 8.10 (d, 1H), 8.18 (d, 1H), 8.7 (d, 2H),9.1 (d, 2H), LC/MS m/z 414 rt=4.43 min.

EXAMPLE 19 Preparation of4-(5-bromoindolin-1-yl)-7-(4-pyridylmethoxy)furo[2,3-d]pyridazine

4,7-Dichloro[2,3-d]furopyridazine from step 4 of Example 9 (95 mg, 0.50mmol) and 5-bromoindoline (100 mg, 0.50 mmol) were refluxed in 60 mL ofabsolute ethanol at 95° C. for 2 hrs. The reaction mixture was allowedto cool to room temperature and the precipitate that formed was filteredand washed with isopropyl alcohol, 4.0 N KOH, H₂O, and hexane, and thendried. The intermediate of about 95% purity (rt=4.72, (M+H)⁺350) and wasused in the next step without further purification. 4-Pyridylcarbinol(28 mg, 0.26 mmol) and sodium hydride (60%, 50 mg, 1.25 mmol) werestirred in 20 mL of anhydrous tetrahydrofuran at 0° C. under Argon for20 min. and then 44 mg of the above intermediate (0.13 mmol) was added.The reaction was stirred at 0° C. for 2 hrs and the temperature wasallowed to rise to room temperature. The mixture was stirred for another12 hrs and the solvent was evaporated under reduced pressure. The solidthat was obtained was dissolved in 50 mL of dichloromethane and washedwith K₂CO₃ solution and H₂O. The organic layer was separated, dried(MgSO₄), and evaporated under reduced pressure. The crude product waspurified by preparative TLC (R_(f)=0.3) on silica gel usingdichloromethane/methanol (95:5) as the eluent. The structure of the puretitle compound was confirmed by LC/MS and NMR: ¹H NMR (CDCl₃) δ 3.20 (m,2H), 4.30˜4.50 (m, 2H), 5.60 (s, 2H), 6.9˜8.0 (m, 7 H), 8.60 (m, 2H);LC/MS (M+H)⁺423 rt=4.49 min.

EXAMPLE 20 Preparation of4-(4-methoxyphenylamino)-7-(2-methylaminocarbonyl-4-pyridylmethoxy)furo-[2,3-d]pyridazine

To a suspension of 4,7-Dichloro[2,3-d]furopyridazine from step 4 ofExample 9 (400 mg, 2.12 mmol, 1 equiv) and p-anisidine (p-MeOC₆H₄NH₂)(260 mg, 2.12 mmol; 1 equiv) in DME (5 mL) was added water (1 mL). Theresulting solution was heated at 50° C. for 48 h. After cooling to rt,the brown precipitate was removed by filtration and the filtrate wasconcentrated in vacuo to afford the crude product as a brown solid.Trituration of the brown solid with CH₂Cl₂ furnished 292 mg (50%) of theintermediate 4-(4-methoxyphenylamino)-7chlorofuro-[2,3-d]pyridazinewhich was confirmed by LC/MS and NMR. A suspension of this intermediate(292 mg, 1.06 mmol, 1 equiv), (2-methylaminocarbonyl-4-pyridyl)methanol(Intermediate H, 529 mg, 3.18 mmol, 3 equiv) and 18-crown-6 (42 mg, 0.16mmol, 15 mol %) in toluene (4 mL) was stirred at rt for 20 min. KOH (178mg, 3.18 mmol, 3 equiv) was then added and the reaction mixture washeated to 80° C. for 36 h. After cooling to rt, water (10 mL) was addedand the mixture was stirred vigorously until a fine white suspension wasformed. The suspension was filtered and washed with water and diethylether to provide 125 mg (29%) of the desired product as a light yellowsolid: (M+H)⁺=406; R_(f)=0.50 (100% EtOAc).

EXAMPLE 21 Preparation of4-(4-methoxyphenylamino)-7-(4-pyridylmethoxy)furo-[2,3-d]pyridazine

The procedure used for the preparation of Example 20 was used to preparethe title compound by substituting 4-pyridylmethanol for(2-methylaminocarbonyl-4-pyridyl)methanol. The pure product was isolatedby chromatography on a flash column: (M+H)⁺349; R_(f)=0.3 (95:5CH₂Cl₂/CH₃OH).

EXAMPLE 22 Preparation of4-(4-methoxyphenylamino)-7-(2-aminocarbonyl4-pyridylmethoxy)furo-[2,3-d]pyridazine

The procedure used for the preparation of Example 16 was used to preparethe title compound by substituting the product of Example 21 for theproduct from Example 9. The reaction was conducted with 250 mg of thestarting material and proportional amounts of solvents and reagents. Thecrude product was purified by HPLC on a 75×30 mm C18 column and a lineargradient elution from 10 to 100% acetonitrile in water with 0.1%trifluoroacetic acid at 10 ml/min. over 10 min. to yield 16 mg of thetitle compound as a yellow solid: HPLC (50×4.6 mm YMC CombiScreen® C18column, linear gradient 10 to 100% acetonitrile in water with 0.1%trifluoroacetic acid at 3 ml/min. over 5 min., UV detection at 254 nm)1.98 min. peak; MS ES 392 (M+H)⁺.

EXAMPLES 23-80 Preparation of Invention Compounds by Methods A-1,A-2 andA-3

Method A-1

Equal equivalents of dichloride (1) and M—NH₂ are refluxed in theappropriate amount of absolute ethanol at 95° C. for 2 hrs. The reactionmixture is allowed to cool to room temperature and the precipitate (2)that forms is filtered and washed sequentially with isopropyl alcohol,4.0 N KOH, H₂O, and hexane, and then dried. The filtrate (2) is thenreacted with 1.2 equivalent of Q—NH₂ in an appropriate amount of n-butylalcohol at 150° C. for 10 hrs. The reaction is cooled to roomtemperature before the solvent is evaporated under reduced pressure. Theresidue is treated with aqueous 4.0 N KOH solution and extracted withdichloromethane. The organic layer is dried (MgSO₄) and evaporated. Thecrude product (3) is purified by preparative thin layer chromatography(TLC) or flash chromatography on silica gel usingdichloromethane/methanol (95:5) as the eluent. Final product isconfirmed by LC/MS and/or NMR. The invention compounds of Examples23-25, 48, and 76-80 as shown in the below table were prepared by methodA-1.

Method A-2

One equivalent of dichloride (1) and 2.2 equivalent of M—NH₂ arerefluxed in an appropriate amount of n-butanol at 150° C. for 10 hrs.The reaction mixture is allowed to cool to room temperature and theprecipitate (4) that forms is filtered and washed sequentially withisopropyl alcohol, 4.0 N KOH, H₂O, and hexane, and then dried. The crudeproduct (4) is purified by preparative TLC or flash chromatography onsilica gel using dichloromethane/methanol (95:5) as the eluent. Finalproduct is confirmed by LC/MS and/or NMR. The invention compounds ofExamples 26-33 and 75 as shown in the below table were prepared bymethod A-2.

Method A-3

One equivalent of dichloride (1) and one equivalent of M—NH₂ aresuspended in DME (0.3M) and water is added until a solution was formed.The reaction mixture is heated to 65° C. for 48 h. After cooling to rt,the resulting precipitate is filtered and washed with DME to provide theintermediate product (2) which is confirmed by LC/MS and NMR. In someinstances, intermediate (2) is further purified by preparative TLC orwashed with other solvents. A suspension of (2) (1 equiv), carbinol (3)(3 equiv), and 18-crown-6 (10 mol %) in toluene (0.3M) is stirred at rtfor 10 min. KOH (3 equiv) is then added and the reaction mixture isheated to 80° C. for 24 h. After cooling to rt, water is added and themixture is stirred vigorously until a suspension is formed. Thesuspension is filtered and washed with water to provide the desiredproduct (4). Preparative TLC and/or washing with other solvents is usedto further purify final products in some examples. The final productsare assigned by LC/MS and NMR spectroscopy. Final product is confirmedby LC/MS and/or NMR. The invention compounds of Examples 34-47, 49-74,and 81-82D as shown in the below table were prepared by method A-3.

Compounds that were Prepared by Parallel Methods A-1, A-2 or A-3

Ex. # X Y MNH NHQ or QQ Method Characterization* 23 S CH

A-1 m/z = 410 rt = 4.21 min.^(a) 24 S CH

A-1 m/z = 414 rt = 4.43 min.^(a) 25 O CH

A-1^(d) (M + H)⁺ 423 rt = 4.49 min.^(a) 26 S CH

A-2 (M + H)⁺ 401 rt = 2.01 min.^(a) 27 S CH

A-2 (M + H)⁺ 399 rt = 2.27 min.^(a) 28 O CH

A-2 (M + H)⁺ 417 rt = 2.47 min.^(a) 29 O CH

A-2 (M + H)⁺ 385 rt = 1.75 min.^(a) 30 O CH

A-2 (M + H)⁺ 383 rt = 1.83 min.^(a) 31 N N

A-2 (M + H)⁺ 385 rt = 1.62 min.^(a) 32 N N

A-2 (M + H)⁺ 383 rt = 1.88 min.^(a) 33 N N

A-2 (M + H)⁺ 417 rt = 2.47 min.^(a) 34 O CH

A-3 (M + H)⁺ 406 R_(f) = 0.50 (100% EtOAc) 35 O CH

A-3 (M + H)⁺ 410 R_(f) = 0.51 (100% EtOAc) 36 O CH

A-3 (M + H)⁺ 428 R_(f) = 0.55 (100% EtOAc) 37 O CH

A-3 (M + H)⁺ 394 R_(f) = 0.57 (100% EtOAc) 38 O CH

A-3 (M + H)⁺ 455 R_(f) = 0.56 (100% EtOAc) 39 O CH

A-3 (M + H)⁺ 390 R_(f) = 0.53 (100% EtOAc) 40 O CH

A-3 (M + H)⁺ 390 R_(f) = 0.68 (100% EtOAc) 41 O CH

A-3 (M + H)⁺ 419 R_(f) = 0.12 (3:2 CH₂Cl₂/EtOAc) 42 O CH

A-3 (M + H)⁺ 444 R_(f) = 0.60 (100% EtOAc) 43 O CH

A-3 (M + H)⁺ 460 R_(f) = 0.57 (100% EtOAc) 44 O CH

A-3 (M + H)⁺ 440 R_(f) = 0.43 (100% EtOAc) 45 O CH

A-3 (M + H)⁺ 447 R_(f) = 0.07 (100% EtOAc) 46 O CH

A-3 (M + H)⁺ 461 R_(f) = 0.38 (100% EtOAc) 47 O CH

A-3 (M + H)⁺ 412 R_(f) = 0.43 (100% EtOAc) 48 O CH

A-1 (M + H)⁺ 394 R_(f) = 0.37 (100% EtOAc) 49 O CH

A-3 (M + H)⁺ 416 R_(f) = 0.64 (100% EtOAc) 50 O CH

A-3 (M + H)⁺ 406 R_(f) = 0.55 (100% EtOAc) 51 O CH

A-3 (M + H)⁺ 406 R_(f) = 0.52 (100% EtOAc). 52 O CH

A-3 (M + H)⁺ 420 R_(f) = 0.37 (4:1 EtOAc/Hex). 53 O CH

A-3 (M + H)⁺ 444 R_(f) = 0.47 (100% EtOAc). 54 O CH

A-3 (M + H)⁺ 404 R_(f) = 0.49 (100% EtOAc). 55 O CH

A-3 (M + H)⁺ 416 R_(f) = 0.23 (100% EtOAc). 14 O CH

A-3 (M + H)⁺ 410 rt = 2.38 min. 56 O CH

A-3 (M + H)⁺ 349 R_(f) = 0.3 (95:5 CH₂Cl₂/CH₃OH) 57 O CH

A-3 (M + H)⁺ 392 R_(f) = 0.43 (4:1 EtOAc/CH₂Cl₂) 58 O CH

A-3 (M + H)⁺ 335 R_(f) = 0.37 (4/1 EtOAc/CH₂Cl₂) 59 O CH

A-3 (M + H)⁺ 376 R_(f) = 0.32 (4/1 EtOAc/Hex) 60 O CH

A-3 (M + H)⁺ 420 R_(f) = 0.43 (100% EtOAc). 61 O CH

A-3 (M + H)⁺ 466 R_(f) = 0.25 (100% EtOAc). 62 O CH

A-3 (M + H)⁺ 447 R_(f) = 0.11 (4:1 EtOAc/Hex) 63^(c) O CH

A-3 (M + H)⁺ 435 R_(f) = 0.35 (100% EtOAc) 64 O CH

A-3 (M + H)⁺ 383 rt = 1.77 min.^(b) 65 O CH

A-3^(e) (M + H)⁺ 418 R_(f) = 0.50 (100% EtOAc) 66 S CH

A-3^(e) (M + H)⁺ 434 R_(f) = 0.50 (100% EtOAc) 67 S CH

A-3 (M + H)⁺ 410 rt = 2.04 min.^(b) 68 S CH

A-3 (M + H)⁺ 406 rt = 2.36 min.^(b) 69 S CH

A-3 (M + H)⁺ 422 rt = 2.31 min.^(b) 70 S CH

A-3 (M + H)⁺ 476 rt = 2.72 min.^(b) 71 S CH

A-3 (M + H)⁺ 460 rt = 2.39 min.^(b) 72 S CH

A-3 (M + H)⁺ 472 rt = 2.53 min.^(b) 73 S CH

A-3 (M + H)⁺ 432 rt = 2.63 min.^(b) 74 S CH

A-3 (M + H)⁺ 436 rt = 2.26 min.^(b) 75 S CH

A-2 (M + H)⁺ 433 rt = 2.61 min.^(a) 76 S CH

A-1 (M + H)⁺ 455 rt = 3.43 min.^(a) 77 S CH

A-1 (M + H)⁺ 432 rt = 4.05 min.^(a) 78 S CH

A-1 (M + H)⁺ 404 rt = 3.08 min.^(a) 79 S CH

A-1 (M + H)⁺ 408 rt = 3.07 min.^(a) 80 S CH

A-1 (M + H)⁺ 466 rt = 2.86 min.^(a) 81 O CH

A-3 (M + H)⁺ 424 R_(f) = 0.38 (100% EtOAc). 82A O CH

A-3 (M + H)⁺ 467 R_(f) = 0.19 (1:1 EtOAc/CH₃OH). 82B O CH

A-3 (M + H)⁺ 436 R_(f) = 0.78 (100% EtOAc) 82C O CH

A-3^(f) (M + H)⁺ 440 R_(f) = 0.35 (100% EtOAc) 82D O CH

A-3 (M + H)⁺ 424 R_(f) = 0.70 (100% EtOAc) *All compounds in this tablecan be characterized by HPLC - positive ion electrospray massspectroscopy (HPLC ES-MS, conditions as below). In addition some of thecompounds were characterized by TLC on silica gel plates and the R_(f)values and solvents are shown. HPLC retention times are given for otherexamples in this table; ^(a)HPLC-electrospray mass spectra (HPLC ES-MS)were obtained using a Hewlett-Packard 1100 HPLC equipped with aquaternary pump, a variable wavelength detector, a YMC Pro C18 2.0 mm ×23 mm column, and a Finnigan LCQ ion trap mass spectrometer withelectrospray ionization. Gradient elution from 90% A to 95% B over 4minutes was used on the HPLC. Buffer A was 98% water, 2% Acetonitrileand 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA.Spectra were scanned from # 140-1200 amu using a variable ion timeaccording to the number of ions in the source; ^(b)An HPLC assay with UVpeak detection was run in addition to the HPLC ES-MS experiment and theconditions are: 50 × 4.6 mm YMC CombiScreen ® C18 column, lineargradient 10 to 100% acetonitrile in a water with 0.1% trifluoroaceticacid at 3 ml/min. over 5 min., UV detection at 254 nm; ^(c)The productwas purified by RP-HPLC on a C18 column using a water/acetonitrilegradient with added trifluoroacetic acid such that the trifluoroacetatesalt was isolated by evaporation of the pure product;^(d)4-pyridylmethanol, as indicated, was used in step 2 of method A-1rather than an amine; ^(e)For preparation of5-amino-2,3-dihydrobenzofurane see Mitchell, H.; Leblanc, Y. J. Org.Chem. 1994, 59, 682-687. ^(f)The reference to make the known TBSprotected alcohol intermediate is: Parsons, A. F.; Pettifer, R. M. J.Chem. Soc. Perkin Trans. 1, 1998, 651.

The deprotection of

was accomplished in the following manner:

Three equiv of a 1.0 Molar solution of TBAF in THF was added to asolution of the protected alcohol in THF (0.05 Molar) at rt. Thereaction mixture was allowed to stir at rt for 1 h and was quenched withwater followed by extraction with EtOAc.

EXAMPLES 83-92 Preparation of Isoquinolines by Method B-1

Method B-1

Dibromoisoquinoline (5, 29 mg, 0.1 mmol) Example 1, step 1, and M—NH₂(0.2 mmol) in 8-mL vial were heated in 1 mL of n-butanol at 90° C. for36 hrs. The mixture was cooled to room temperature and the solvent wasevaporated under reduced pressure. 4-Mercaptopyridine (23 mg, 0.2 mmol)and cesium carbonate (67 mg, 0.2 mmol) were added to the vial. Themixture was heated at 180° C. for 1 hr and was allowed to cool to roomtemperature. Methanol (2 mL) was added to the vial and the mixture wassonicated for 10 min and filtered. The methanol solution of reactionmixture was collected and evaporated under reduced pressure. Theformation of product was confirmed by LC/MS. The invention compounds ofExamples 83-92 as shown in the below table were prepared by method B-1.

Compounds that were Prepared by Method B-1

Example # MNH Characterization* 83

(M + H)⁺ 412 rt = 3.46 min. 84

(M + H)⁺ 388 rt = 2.89 min. 85

(M + H)⁺ 364 rt = 3.41 min. 86

(M + H)⁺ 346 rt = 1.83 min. 87

(M + H)⁺ 401 rt = 2.52 min. 88

(M + H)⁺ 370 rt = 3.17 min. 89

(M + H)⁺ 387 rt = 3.02 min. 90

(M + H)⁺ 453 rt = 3.39 min. 91

(M + H)⁺ 437 rt = 3.33 min. 92

(M + H)⁺ 401 rt = 2.52 min. *HPLC - electrospray mass spectra (HPLCES-MS) were obtained using a Hewlett-Packard 1100 HPLC equipped with aquaternary pump, a variable wavelength detector, a YMC Pro C18 2.0 mm ×23 mm column, and a Finnigan LCQ ion trap mass spectrometer withelectrospray ionization. Gradient elution from 90% A to 95% B over 4minutes was used on the HPLC. Buffer A was 98% water, 2% Acetonitrileand 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA.Spectra were scanned from # 140-1200 amu using a variable ion timeaccording to the number of ions in the source.

EXAMPLES 93-105 Preparation of Novel Phthalazine Invention Compounds byParallel Synthesis

Method A-1 or A-2, as indicated, were used to prepare the novelphthalimide invention compounds 93-105 from 1,4-dichlorophthalazine (forpreparation see Novartis patent WO98/35958, 11.02.98) rather than thedichloroheterocyclopyridazines together with the appropriate bicyclicand substituted anilines.

Novel Phthalazines that were Prepared by Methods A-1 or A-2

Example # MNH QNH Method Characterization 93

A-2 (M + H)⁺ 427 rt = 3.13 min. 94

A-2 (M + H)⁺ 395 rt = 2.52 min. 95

A-1 (M + H)⁺ 387 rt = 2.77 min. 96

A-1 (M + H)⁺ 388 rt = 2.51 min. 97

A-1 (M + H)⁺ 474 rt = 3.67 min. 98

A-1 (M + H)⁺ 450 rt = 3.54 min. 99

A-1 (M + H)⁺ 453 rt = 2.70 min. 100

A-1 (M + H)⁺ 455 rt = 2.58 min. 101

A-1 (M + H)⁺ 448 rt = 3.02 min. 102

A-1 (M + H)⁺ 412 rt = 3.27 min. 103

A-1 (M + H)⁺ 400 rt = 2.79 min. 104

A-1 (M + H)⁺ 402 rt = 2.96 min. 105

A-1 (M + H)⁺ 404 rt = 3.03 min. * HPLC - electrospray mass spectra (HPLCES-MS) were obtained using a Hewlett-Packard 1100 HPLC equipped with aquaternary pump, a variable wavelength detector, a YMC Pro C18 2.0 mm ×23 mm column, and a Finnigan LCQ ion trap mass spectrometer withelectrospray ionization. Gradient elution from 90% A to 95% B over 4minutes was used on the HPLC. Buffer A was 98% water, 2% Acetonitrileand 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA.Spectra were scanned from # 140-1200 amu using a variable ion timeaccording to the number of ions in the source.

EXAMPLES 106-114 Preparation of Salts of Example 14

The product of Example 14 (1.50 g, 3.66 mmol) was stirred as a slurry inmethanol (20 ml) as a solution of toluenesulfonic acid hydrate (0.701 g,3.67 mmol) in methanol (5 ml plus 5 ml rinse) was added quicklydropwise. All materials dissolved over 5 min to yield a yellow solution.Anhydrous ether (30 ml) was added and stirring was continued for 5minutes until solid began to precipitate. The resultant mixture waschilled with stirring in an ice/water bath for 45 minutes and then thesolid title product (Example 104) was collected by filtration, washedwith ether and dried at 55° C. in a vacuum oven until NMR analysisshowed a lack of solvents (1.5 hours). Other compounds were prepared ina similar way by using a variety of acids rather than toluenesulfonicacid. Scale up and use of less methanol in the first step generally ledto quicker precipitation of salts and a variety of solvents were usedrather than ether, as indicated, to help crystalize the individualsalts. In some cases the methanol was first removed by evaporation invacuo. Final drying took between 1.5 hours and several days, dependingon the quantity of material and the specific specific acid used.

Salts of Example 14 that were Prepared Scale: Characterization Example #Acid Used (14 used, g) Solvent Added (melting point, ° C.) 106

1.5 Ether 167-168 with decomposition 107

0.7 Ether 157-159 108

0.6 Ether 180-182 with decomposition 109

0.7 Ether 153-154 110 (HCl)₂ * in Ether 1.5 Ether 128-131 withdecomposition 111 HBr 0.7 Most MeOH evaporated, then acetone/benzene137-139 with decomposition 112 H₂SO₄ 0.6 Most MeOH evaporated, thenacetone/ether 177-179 with decomposition 113 HNO₃ 0.5 Ether 135(decomposed) melted 150-152 114

0.5 Ether, Prolonged drying, Hygroscopic 123-128 115

4.5 Ether 148-149 * The disalt with HCl was isolated rather than the 1:1salt. This occurred even if less than 2 equivalents of acid were used.

Biological Protocols and In vitro Test Data KDR Assay

The cytosolic kinase domain of KDR kinase was expressed as a 6His fusionprotein in Sf9 insect cells. The KDR kinase domain fusion protein waspurified over a Ni++ chelating column. Ninety-six well ELISA plates werecoated with 5 μg poly(Glu4;Tyr1) (Sigma Chemical Co., St Louis, Mo.) in100 μl HEPES buffer (20 mM HEPES, pH 7.5, 150 mM NaCl, 0.02% Thimerosal)at 4° overnight. Before use, the plate was washed with HEPES, NaClbuffer and the plates were blocked with 1% BSA, 0.1% Tween 20 in HEPES,NaCl buffer.

Test compounds were serially diluted in 100% DMSO from 4 mM to 0.12 μMin half-log dilutions. These dilutions were further diluted twenty foldin H2O to obtain compound solutions in 5% DMSO. Following loading of theassay plate with 85 μl of assay buffer (20 mM HEPES, pH 7.5, 100 mM KCl,10 mM MgCl₂, 3 mM MnCl₂, 0.05% glycerol, 0.005% Triton X-100, 1mM-mercaptoethanol, with or without 3.3 μM ATP), 5 μl of the dilutedcompounds were added to a final assay volume of 100 μl. Finalconcentrations were between 10 μM, and 0.3 nM in 0.25% DMSO. The assaywas initiated by the addition of 10 μl (30 ng) of KDR kinase domain.

The assay was incubated with test compound or vehicle alone with gentleagitation at room temperature for 60 minutes. The wells were washed andphosphotyrosines (PY) were probed with an anti-phosphotyrosine (PY), mAbclone 4G10 (Upstate Biotechnology, Lake Placid, N.Y.). PY/anti-PYcomplexes were detected with an anti-mouse IgG/HRP conjugate (AmershamInternational plc, Buckinghamshire, England). Phosphotyrosine wasquantitated by incubating with 100 μl 3,3′,5,5′tetramethylbenzidinesolution (Kirkegaard and Perry, TMB Microwell 1 Component peroxidasesubstrate). Color development was arrested by the addition of 100 μl 1%HCl-based stop solution (Kirkegaard and Perry, TMB 1 Component StopSolution).

Optical densities were determined spectrophotometrically at 450 nm in a96-well plate reader, SpectraMax 250 (Molecular Devices). Background (noATP in assay) OD values were subtracted from all ODs and the percentinhibition was calculated according to the equation:${\% \quad {Inhibition}} = \frac{\left( {{{OD}\left( {{vehicle}\quad {control}} \right)} - {{OD}\left( {{with}\quad {compound}} \right)}} \right) \times 100}{{{OD}\left( {{vehicle}\quad {control}} \right)} - {{OD}\left( {{no}\quad {ATP}\quad {added}} \right)}}$

The IC₅₀ values were determined with a least squares analysis programusing compound concentration versus percent inhibition. Compounds thathave IC₅₀≦100 nM in this assay include those of Examples 1, 2, 4, 6, 8,9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 22, 23, 24, 34, 37, 38, 39,40, 42, 43, 44, 47, 49, 51, 52, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66,68, 69, 70, 71, 72, 73, 74, 75, 78, 82B, 82C, 82D, 85, 88, 93, 96, 97,98, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, and 112.Compounds that have IC₅₀ values between 100 nM and 1,000 nM includethose of examples 3, 5, 7, 21, 27, 28, 35, 36, 45, 46, 48, 50, 55, 58,61, 64, 67, 76, 79, 82A, 89, 95, 99, and 100. Those that have measuredIC₅₀ values>1,000 nM include those of examples 26, 29, 30, 31, 32, 33,41, 77, 80, 81, and 94. Example numbers not in this list may be assumedto be weakly active, with IC₅₀ values greater than 1 μM.

Cell Mechanistic Assay-Inhibition of 3T3 KDR Phosphorylation

NIH3T3 cells expressing the full length KDR receptor were grown in DMEM(Life Technologies, Inc., Grand Island, N.Y.) supplemented with 10%newborn calf serum, low glucose, 25 mM/L sodium pyruvate, pyridoxinehydrochloride and 0.2 mg/ml of G418 (Life Technologies Inc., GrandIsland, N.Y.). The cells were maintained in collagen I-coated T75 flasks(Becton Dickinson Labware, Bedford, Mass.) in a humidified 5% CO2atmosphere at 37° C.

Fifteen thousand cells were plated into each well of a collagen I-coated96-well plate in the DMEM growth medium. Six hours later, the cells werewashed and the medium was replaced with DMEM without serum. Afterovernight culture to quiesce the cells, the medium was replaced byDulbecco's phosphate-buffered saline (Life Technologies Inc., GrandIsland, N.Y.) with 0.1% bovine albumin (Sigma Chemical Co., St Louis,Mo.). After adding various concentrations (0-300 nM) of test compoundsto the cells in 1% final concentration of DMSO, the cells were incubatedat room temperature for 30 minutes. The cells were then treated withVEGF (30 ng/ml) for 10 minutes at room temperature. Following VEGFstimulation, the buffer was removed and the cells were lysed by additionof 150 μl of extraction buffer (50 mM Tris, pH 7.8, supplemented with10% glycerol, 50 mM BGP, 2 mM EDTA, 10 mM NaF, 0.5 mM NaVO4, and 0.3%TX-100) at 4° C. for 30 minutes.

To assess receptor phosphorylation, 100 microliters of each cell lysatewas added to the wells of an ELISA plate precoated with 300 ng ofantibody C20 (Santa Cruz Biotechnology, Inc., Santa Cruz, Calif.).Following a 60-minute incubation, the plate was washed and bound KDR wasprobed for phosphotyrosine using an anti-phosphotyrosine mAb clone 4G10(Upstate Biotechnology, Lake Placid, N.Y.). The plate was washed andwells were incubated with anti-mouse IgG/HRP conjugate (AmershamInternational plc, Buckinghamshire, England) for 60 minutes. Wells werewashed and phosphotyrosine was quantitated by addition of 100 μl perwell of 3,3′,5,5′tetramethylbenzidine (Kirkegaard and Perry, TMBMicrowell 1 Component peroxidase substrate) solution. Color developmentwas arrested by the addition of 100 μl 1% HCl based stop solution(Kirkegaard and Perry, TMB 1 Component Stop Solution).

Optical densities (OD) were determined spectrophotometrically at 450 nmin a 96-well plate reader (SpectraMax 250, Molecular Devices).Background (no VEGF added) OD values were subtracted from all ODs andpercent inhibition was calculated according to the equation:${\% \quad {Inhibition}} = \frac{\left( {{{OD}\left( {{VEGF}\quad {control}} \right)} - {{OD}\left( {{with}\quad {test}\quad {compound}} \right)}} \right) \times 100}{{{OD}\left( {{VEGF}\quad {control}} \right)} - {{OD}\left( {{no}\quad {VEGF}\quad {added}} \right)}}$

IC₅₀s were determined on some of the exemplary materials with a leastsquares analysis program using compound concentration versus percentinhibition. Compounds that have IC₅₀≦20 nM in this assay include thoseof Examples 2, 6, 10, 11, 14, 23, 96, 101, 102, 103, 104, 105. Compoundsthat have IC₅₀ values between 20 nM and 50 nM include those of examples1, 4, 8, 9, 12, 13, 17, 24, 93, 98. Compounds that have IC₅₀ valuesbetween 50 nM and 400 nM include those of examples 97, 99, and 100.

Matrigel® Angiogenesis Model Preparation of Matrigel Plugs and In vivoPhase

Matrigel® (Collaborative Biomedical Products, Bedford, Mass.) is abasement membrane extract from a murine tumor composed primarily oflaminin, collagen IV and heparan sulfate proteoglycan. It is provided asa sterile liquid at 4° C., but rapidly forms a solid gel at 37° C.

Liquid Matrigel at 4° C. was mixed with SK-MEL2 human tumor cells thatwere transfected with a plasmid containing the murine VEGF gene with aselectable marker. Tumor cells were grown in vitro under selection andcells were mixed with cold liquid Matrigel at a ratio of 2×10⁶ per 0.5ml. One half milliliter was implanted subcutaneously near the abdominalmidline using a 25 gauge needle. Test compounds were dosed as solutionsin Ethanol/Cremaphor EL/saline (12.5%:12.5%:75%) at 30, 100, and 300mg/kg po once daily starting on the day of implantation. Mice wereeuthanized 12 days post-implantation and the Matrigel pellets wereharvested for analysis of hemoglobin content.

Hemoglobin Assay

The Matrigel pellets were placed in 4 volumes (w/v) of 4° C. LysisBuffer (20 mM Tris pH 7.5, 1 mM EGTA, 1 mM EDTA, 1% Triton X-100 [EMScience, Gibbstown, N.J.], and complete, EDTA-free protease inhibitorcocktail [Mannheim, Germany]), and homogenized at 4° C. Homogenates wereincubated on ice for 30 minutes with shaking and centrifuged at 14K×gfor 30 minutes at 4° C. Supernatants were transferred to chilledmicrofuge tubes and stored at 4° C. for hemoglobin assay.

Mouse hemoglobin (Sigma Chemical Co., St. Louis, Mo.) was suspended inautoclaved water (BioWhittaker, Inc, Walkersville, Md.) at 5 mg/ml. Astandard curve was generated from 500 micrograms/ml to 30 micrograms/mlin Lysis Buffer (see above). Standard curve and lysate samples wereadded at 5 microliters/well in duplicate to a polystyrene 96-well plate.Using the Sigma Plasma Hemoglobin Kit (Sigma Chemical Co., St. Louis,Mo.), TMB substrate was reconstituted in 50 mls room temperature aceticacid solution. One hundred microliters of substrate was added to eachwell, followed by 100 microliters/well of Hydrogen Peroxide Solution atroom temperature. The plate was incubated at room temperature for 10minutes.

Optical densities were determined spectrophotometrically at 600 nm in a96-well plate reader, SpectraMax 250 Microplate Spectrophotometer System(Molecular Devices, Sunnyvale, Calif.). Background Lysis Buffer readingswere subtracted from all wells.

Total sample hemoglobin content was calculated according to thefollowing equation:

Total Hemoglobin=(Sample Lysate Volume)×(Hemoglobin Concentration)

The average Total Hemoglobin of Matrigel samples without cells wassubtracted from each Total Hemoglobin Matrigel sample with cells.Percent inhibition was calculated according to the following equation:${\% \quad {Inhibition}} = \frac{\text{(Average~~Total~~Hemoglobin~~Drug-Treated~~Tumor~~Lysates)} \times \quad 100}{\text{(Average~~Total~~Hemoglobin~~Non-Treated~~Tumor~~Lysates)}}$

Example 8 showed significant activity in this assay at 100 and 300 mg/kgpo sid with >60% inhibition of total hemoglobin content of the Matrigelsamples from the dosed animals vs. those from vehicle control animals.The other examplary materials were not tested in this model.

Other embodiments of the invention will be apparent to the skilled inthe art from a consideration of this specification or practice of theinvention disclosed herein. It is intended that the specification andexamples be considered as exemplary only, with the true scope and spiritof the invention being indicated by the following claims.

We claim:
 1. A compound having the generalized structural formula

wherein R¹ and R² together form a bridge containing two T² moieties andone T³ moiety, said bridge, taken together with the ring to which it isattached, forming a bicyclic of structure

 wherein each T² independently represents N, CH, or CG¹; and T³represents S, O, CR⁴G¹, C(R⁴)₂, or NR³;  and wherein G¹ is a substituentindependently selected from the group consisting of —N(R⁶)₂; —NR³COR⁶;halogen; alkyl; cycloalkyl; lower alkenyl; lower cycloalkenyl;halogen-substituted alkyl; amino-substituted alkyl; N-loweralkylamino-substituted alkyl; N,N-di-lower alkylamino-substituted alkyl;N-lower alkanoylamino-substituted alkyl; hydroxy-substituted alkyl;cyano-substituted alkyl; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted saturated heterocyclylalkyl;optionally substituted partially unsaturated heterocyclyl; optionallysubstituted partially unsaturated heterocyclylalkyl; —OCO₂R³; optionallysubstituted heteroarylalkyl; optionally substituted heteroaryloxy;—S(O)_(p)(optionally substituted heteroaryl); optionally substitutedheteroarylalkyloxy; —S(O)_(p)(optionally substituted heteroarylalkyl);—CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂ R³ is H or lower alkyl; R⁶ isindependently selected from the group consisting of H; alkyl;cycloalkyl; optionally substituted aryl; and optionally substituted aryllower alkyl; lower alkyl-N(R³)₂; and lower alkyl-OH; R⁴ is H, halogen,or lower alkyl; p is 0, 1, or 2; X is selected from the group consistingof O, S, and NR³; Y is selected from the group consisting of loweralkylene; —CH₂—O—; —CH₂—S—; —CH₂—NH—; —O—; —S—; —NH—; —(CR⁴₂)_(n)—S(O)_(p)-(5-membered heteroaryl)-(CR⁴ ₂)_(s)—; —(CR⁴₂)_(n)—C(G²)(R⁴)—(CR⁴ ₂)_(s)—;  wherein n and s are each independently 0or an integer of 1-2; and G² is selected from the group consisting of—CN, —CO₂R³, —CON(R⁶)₂, and —CH₂N(R⁶)₂; —O—CH₂—; —S(O)—; —S(O)₂—;—SCH₂—; —S(O)CH₂—; —S(O)₂CH₂—; —CH₂S(O)—; and —CH₂S(O)₂— Z is N; q is 0,1, or 2; G³ is a monovalent or bivalent moiety selected from the groupconsisting of: lower alkyl; —NR³COR⁶; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶; —OCOR⁶;—COR⁶; —CO₂R⁶; —CH₂OR³; —CON(R⁶)₂; —S(O)₂N(R⁶)₂; —NO₂; —CN; optionallysubstituted aryl; optionally substituted heteroaryl; optionallysubstituted saturated heterocyclyl; optionally substituted partiallyunsaturated heterocyclyl; optionally substituted heteroarylalkyl;optionally substituted heteroaryloxy; —S(O)_(p)(optionally substitutedheteroaryl); optionally substituted heteroarylalkyloxy;—S(O)_(p)(optionally substituted heteroarylalkyl); —OCON(R⁶)₂;—NR³CO₂R⁶; —NR³CON(R⁶)₂; and bivalent bridge of structure T²═T²—T³ wherein each T² independently represents N, CH, or CG^(3′); and T³represents S, O, CR⁴G^(3′), C(R⁴)₂, or NR³; wherein G^(3′) representsany of the above-defined moieties G³ which are monovalent; and theterminal T² is bound to L, and T³ is bound to D, forming a 5-memberedfused ring; A and D independently represent N or CH; B and Eindependently represent N or CH; L represents N or CH; and  with theprovisos that a) the total number of N atoms in the ring containing A,B, D, E, and L is 0, 1, 2, or 3; and b) when L represents CH and q=0 orany G³ is a monovalent substituent, at least one of A and D is an Natom; and c) when L represents CH and a G³ is a bivalent bridge ofstructure T²═T²—T³, then A, B, D, and E are also CH; J is a ringselected from the group consisting of aryl; pyridyl; and cycloalkyl; q′represents the number of substituents G⁴ on ring J and is 0, 1, 2, 3, 4,or 5, and G⁴ is a monovalent or bivalent moiety selected from the groupconsisting of —N(R⁶)₂; —NR³COR⁶; halogen; alkyl; cycloalkyl; loweralkenyl; lower cycloalkenyl; halogen-substituted alkyl;amino-substituted alkyl; N-lower alkylamino-substituted alkyl;N,N-di-lower alkylamino-substituted alkyl; N-loweralkanoylamino-substituted alkyl; hydroxy-substituted alkyl;cyano-substituted alkyl; carboxy-substituted alkyl; loweralkoxycarbonyl-substituted alkyl; phenyl loweralkoxycarbonyl-substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; N-lower alkylamino-substituted alkylamino;N,N-di-lower alkylamino-substituted alkylamino; N-loweralkanoylamino-substituted alkylamino; hydroxy-substituted alkylamino;cyano-substituted alkylamino; carboxy-substituted alkylamino; loweralkoxycarbonyl-substituted alkylamino; phenyl-loweralkoxycarbonyl-substituted alkylamino; —OR⁶; —SR⁶; —S(O)R⁶; —S(O)₂R⁶;halogenated lower alkoxy; halogenated lower alkylthio; halogenated loweralkylsulfonyl; —OCOR⁶; —COR⁶; —CO₂R⁶; —CON(R⁶)₂; —CH₂OR³; —NO₂; —CN;amidino; guanidino; sulfo; —B(OH)2; optionally substituted aryl;optionally substituted heteroaryl; optionally substituted saturatedheterocyclyl; optionally substituted partially unsaturated heterocyclyl;—OCO₂R³; optionally substituted heteroarylalkyl; optionally substitutedheteroaryloxy; —S(O)_(p)(optionally substituted heteroaryl); optionallysubstituted heteroarylalkyloxy; —S(O)_(p)(optionally substitutedheteroarylalkyl); —CHO; —OCON(R⁶)₂; —NR³CO₂R⁶; —NR³CON(R⁶)₂; and fusedring-forming bivalent bridges attached to and connecting adjacentpositions of ring J, said bridges having the structures:

 wherein each T² independently represents N, CH, or CG^(4′); T³represents S, O, CR⁴G^(4′), C(R⁴)₂, or NR³; wherein G^(4′) representsany of the above-defined moieties G⁴ which are monovalent; and bindingto ring J is achieved via terminal atoms T² and T³;

 wherein each T² independently represents N, CH, or CG^(4′); whereinG^(4′) represents any of the above-defined moieties G⁴ which aremonovalent; and with the proviso that a maximum of two bridge atoms T²may be N; and binding to ring J is achieved via terminal atoms T²; and

 wherein each T⁴, T⁵, and T⁶ independently represents O, S, CR⁴G^(4′),C(R⁴)₂, or NR³; wherein G^(4′) represents any of the above-definedmoieties G⁴ which are monovalent; and binding to ring J is achieved viaterminal atoms T⁴ or T⁵; with the provisos that: i) when one T⁴ is O, S,or NR³, the other T⁴ is CR⁴G^(4′) or C(R⁴)₂; ii) a bridge comprising T⁵and T⁶ atoms may contain a maximum of two heteroatoms O, S, or N; andiii) in a bridge comprising T⁵ and T⁶ atoms, when one T⁵ group and oneT⁶ group are O atoms, or two T⁶ groups are O atoms, said O atoms areseparated by at least one carbon atom; when G⁴ is an alkyl group locatedon ring J adjacent to the linkage —(CR⁴ ₂)_(p)—, and X is NR³ wherein R³is an alkyl substituent, then G⁴ and the alkyl substituent R³ on X maybe joined to form a bridge of structure —(CH₂)_(p′)— wherein p′ is 2, 3,or 4, with the proviso that the sum of p and p′ is 2, 3, or 4, resultingin formation of a nitrogen-containing ring of 5, 6, or 7 members; andwith the further provisos that: in G¹, G², G³, and G⁴, when two groupsR³ or R⁶ are each alkyl and located on the same N atom they may belinked by a bond, an O, an S, or NR³ to form a N-containing heterocycleof 5-7 ring atoms; when an aryl, heteroaryl, or heterocyclyl ring isoptionally substituted, that ring may bear up to 5 substituents whichare independently selected from the group consisting of amino,mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino,lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl,hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy,halogenated lower alkylthio, lower alkanoyloxy, —CO₂R³, —CHO, —CH₂OR³,—OCO₂R³, —CON(R⁶)₂, —OCON(R⁶)₂, —NR³CON(R⁶)₂, nitro, amidino, guanidino,mercapto, sulfo, and cyano; and when any alkyl group is attached to O,S, or N, and bears a hydroxyl substituent, then said hydroxylsubstituent is separated by at least two carbon atoms from the O, S, orN to which the alkyl group is attached; or a pharmaceutically acceptablesalt thereof.
 2. A compound of claim 1 wherein R¹ and R² together form abridge containing two T² moieties and one T³ moiety, said bridge, takentogether with the ring to which it is attached, forming a bicyclic ofstructure

 wherein each T² independently represents N, CH, or CG¹; and T³represents S, O, CH₂, or NR³; with the proviso that when T³ is O or S,at least one T² is CH or CG¹.
 3. A pharmaceutical composition comprisinga compound of claim 1 and a pharmaceutically acceptable carrier.
 4. Amethod of treating a mammal having a condition characterized by abnormalangiogenesis or hyperpermiability processes, comprising administering tosaid mammal an amount of a compound of claim 1 which is effective totreat said condition.
 5. The method of claim 4, wherein said conditionis tumor growth; retinopathy, including diabetic retinopathy, ischemicretinal-vein occlusion, retinopathy of prematurity, and age-relatedmacular degeneration; rheumatoid arthritis; psoriasis; or a bullousdisorder associated with subepidermal blister formation, includingbullous pemphigoid, erythema multiforme, and dermatitis herpetiformis.6. A compound of claim 1 selected from the group consisting of thefollowing named exemplary compounds: Ex- am- ple No. Chemical Name  8N-(4-chlorophenyl)-7-(4-pyridinylmethoxy)thieno[2,3-d]py-ridazin-4-amine  9N-(4-chlorophenyl)-7-(4-pyridinylmethoxy)furo[2,3-d]py- ridazin-4-amine 10 4-[({4-[(4-chlorophenyl)amino]thieno[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinecarboxamide  114-[({4-[(4-chlorophenyl)amino]thieno[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  144-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  164-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinecarboxamide  17N-(1,3-benzothiazol-6-yl)-N-{4-[(4-chlorophenyl)ami-no]thieno[2,3-d]pyridazin-7-yl}amine  18N-(1,3-benzothiazol-6-yl)-N-[4-(2,3-dihydro-1H-inden-5-yl-amino)thieno[2,3-d]pyridazin-7-yl]amine  194-(5-bromo-2,3-dihydro-1H-indol-1-yl)-7-(4-pyridinyl-methoxy)furo[2,3-d]pyridazine  204-[({4-[(4-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  21N-(4-methoxyphenyl)-7-(4-pyridinylmethoxy)furo[2,3-d]py- ridazin-4-amine 22 4-[({4-[(4-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinecarboxamide  23N⁷-(1,3-benzothiazol-6-yl)-N⁴-(4-chloro-phenyl)thieno[2,3-d]pyridazine-4,7-diamine  24N-(1,3-benzothiazol-6-yl)-N-[4-(2,3-dihydro-1H-inden-5-yl-amino)thieno[2,3-d]pyridazin-7-yl]amine  27N-(1H-indazol-5-yl)-N-[4-(1H-indazol-5-ylamino)thi-eno[2,3-d]pyridazin-7-yl]amine  28N-(1,3-benzothiazol-6-yl)-N-[4-(1,3-benzothiazol-6-yl-amino)furo[2,3-d]pyridazin-7-yl]amine  344-[({4-[(4-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  354-[({4-[(3-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  364-[({4-[(3-chloro-4-fluorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  374-[({4-[(4-fluorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  384-[({4-[(4-bromophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  39N-methyl-4-[({4-[(4-methylphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinecarboxamide  40N-methyl-4-[({4-[(3-methylphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinecarboxamide  42 N-methyl-4-{[(4-{[4-(trifluoromethyl)phenyl]ami- no}furo[2,3-d]pyridazin-7-yl)oxy]me-thyl}-2-pyridinecarboxamide  43N-methyl-4-{[(4-{[4-(trifluoromethoxy)phenyl]ami-no}furo[2,3-d]pyridazin-7-yl)oxy]me- thyl}-2-pyridinecarboxamide  444-[({4-[(3-chloro-4-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  454-({[4-({4-[acetyl(methyl)amino]phenyl}ami-no)furo[2,3-d]pyridazin-7-yl]oxy}methyl)-N-methyl-2-py-ridinecarboxamide  46 N-methyl-4-{[(4-{[4-(4-morpholinyl)phenyl]ami-no}furo[2,3-d]pyridazin-7-yl)oxy]methyl}-2-py- ridinecarboxamide  474-[({4-[(3,4-difluorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  48N-(1,3-benzothiazol-6-yl)-N-{4-[(4-chlorophenyl)ami-no]furo[2,3-d]pyridazin-7-yl}amine  494-({[4-(2,3-dihydro-1H-inden-5-ylamino)furo[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  504-[({4-[(2-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  514-[({4-[(3-methoxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  524-({[4-(1,3-benzodioxol-5-ylamino)furo[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  534-[({4-[(3,4-dichlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  544-[({4-[(3,5-dimethylphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  554-({[4-(1H-indazol-5-ylamino)furo[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  56N-(4-methoxyphenyl)-7-(4-pyridinylmethoxy)furo[2,3-d]py- ridazin-4-amine 57 4-[({4-[(4-hydroxyphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  584-{[7-(4-pyridinylmethoxy)furo[2,3-d]py- ridazin-4-yl]amino}phenol  594-{[(4-anilinofuro[2,3-d]pyridazin-7-yl)oxy]me-thyl}-N-methyl-2-pyridinecarboxamide  604-[({4-[(3-methoxy-4-methylphenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  61N-(4-chlorophenyl)-7-{[2-(4-morpholinylcarbonyl)-4-pyridinyl]methoxy}furo[2,3-d]pyridazin-4-amine  62N-methyl-4-[({4-[(2-methyl-1,3-benzothiazol-5-yl)ami-no]furo[2,3-d]pyridazin-7-yl}oxy)methyl]-2-py- ridinecarboxamide  634-({[4-(1,3-benzothiazol-6-ylamino)furo[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide trifluoroacetate 64 {4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-2-pyridinyl}methanol  654-({[4-(2,3-dihydro-1-benzofuran-5-ylamino)furo[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  664-({[4-(2,3-dihydro-1-benzofuran-5-ylamino)thi-eno[2,3-d]pyridazin-7-yl]oxy}methyl)-N-me- thyl-2-pyridinecarboxamide 67 4-[({4-[(4-fluorophenyl)amino]thieno[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  68N-methyl-4-[({4-[(3-methylphenyl)amino]thi-eno[2,3-d]pyridazin-7-yl}oxy)methyl]-2-py- ridinecarboxamide  694-[({4-[(4-methoxyphenyl)amino]thieno[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  70N-methyl-4-{[(4-{[4-(trifluoromethoxy)phenyl]ami-no}thieno[2,3-d]pyridazin-7-yl)oxy]methyl }-2-py- ridinecarboxamide  71N-methyl-4-{[(4-{[4-(trifluoromethyl)phenyl]ami-no}thieno[2,3-d]pyridazin-7-yl)oxy]methyl}-2-py- ridinecarboxamide  724-[({4-[(4-bromophenyl)amino]thieno[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridinecarboxamide  734-({[4-(2,3-dihydro-1H-inden-5-ylamino)thieno[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  744-({[4-(1,3-benzodioxol-5-ylamino)thieno[2,3-d]py-ridazin-7-yl]oxy}methyl)-N-methyl-2-pyridinecarboxamide  75N-(1,3-benzothiazol-6-yl)-N-[4-(1,3-benzothiazol-6-yl-amino)thieno[2,3-d]pyridazin-7-yl]amine  76N-(1,3-benzothiazol-6-yl)-N-{4-[(4-bromophenyl)amino]thi-eno[2,3-d]pyridazin-7-yl}amine  78N-(1,3-benzothiazol-6-yl)-N-{4-[(2,4-dimethyl-phenyl)amino]thieno[2,3-d]pyridazin-7-yl}amine  79N-(1,3-benzothiazol-6-yl)-N-{4-[(3-fluoro-4-methyl-phenyl)amino]thieno[2,3-d]pyridazin-7-yl}amine  82A4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-[2-(dimethyl-amino)ethyl]-2-pyridinecarboxamide  82B4-[({4-[(4-chlorophenyl)amino]furo[2 ,3-d]py-ridazin-7-yl}oxy)methyl]-N-cyclo- propyl-2-pyridinecarboxamide  82C4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-(2-hydroxyethyl)-2-py- ridinecarboxamide  82D4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-ethyl-2-pyridinecarboxamide 1064-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide4-methylbenzenesulfonate 1074-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide4-chlorobenzenesulfonate 1084-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamidemethanesulfonate 109 4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamideethanesulfonatesulfonate 1104-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamidedihydrochloride 111 4-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide hydrobromide112 4-[({4-[(4-chlorophenyl)amino]furo [2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide sulfate 1134-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide nitrate 1144-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamide2-hydroxyethanesulfonate 1154-[({4-[(4-chlorophenyl)amino]furo[2,3-d]py-ridazin-7-yl}oxy)methyl]-N-methyl-2-pyridine- carboxamidebenzenesulfonate